METHODS AND COMPOSITIONS FOR THE SELECTIVE REGULATION OF PROTEIN EXPRESSION

Abstract

The present invention relates to a recombinant DNA molecule characterized in that it comprises an mts-rRNA target element comprising a sequence selected from the group consisting of SEQ ID NO: 7, 8 and complements thereof wherein the mts-rRNA target element is operatively linked to a transcribable heterologous polynucleotide molecule.

Description

METHODS AND COMPOSITIONS FOR THE SELECTIVE REGULATION OF PROTEIN EXPRESSION CROSS REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 62 / 195.546, filed on July 22, 2015, which is incorporated herein in its entirety by reference. INCORPORATION OF SEQUENCE LISTS The list of sequences contained in the file named MONS392US_ST25.txt, which is 26.3 kilobytes in size (measured on the MS-Windows operating system), created on June 28, 2016, is presented herein and incorporated herein by reference. BACKGROUND OF THE INVENTION Field of invention The invention relates generally to the fields of agriculture, plant breeding, and molecular biology. More specifically, the invention relates to methods and compositions for selectively regulating protein expression in the male reproductive tissue of transgenic plants and their uses. Description of the related technique A hybrid seed is produced by hybridizing or cross-fertilizing closely related plants and can be grown into hybrid progeny plants that possess a desired combination of traits not found in either of the original plants. Hybrid plants can exhibit superior agronomic characteristics such as improved plant size, yield, nutritional composition, disease resistance, herbicide tolerance, stress tolerance, climate adaptation, and other desirable traits. Efficient hybrid seed production requires that a plant's pollen cannot self-fertilize. A major limitation in hybrid seed production for many crops is the lack of simple, reliable, and inexpensive methods for male sterilizing plants and preventing self-fertilization. RQonnn / i zciz / r / yiai In hybrid seed production, pollen production and dispersal can be prevented in a female parent plant to facilitate cross-pollination rather than self-pollination. This can be achieved, for example, by manually removing pollen-bearing structures (e.g., by manual or mechanical detasseling in maize), using a genetic pollination control method (e.g., using cytoplasmic or nuclear male sterilization technology), using a chemical agent, or any combination thereof. This process can be labor-intensive and therefore costly. In maize, for example, detasseling is typically performed in two stages: mechanical detasseling followed by manual detasseling.The commercial production of hybrid seeds using only chemical gametocides is limited primarily by their general lack of selectivity for gametes and their effect on other parts of the plant. Therefore, methods to improve the efficiency of hybrid seed production are highly desirable. BRIEF DESCRIPTION OF THE INVENTION The invention generally relates to improvements in methods for selectively regulating protein expression in the male reproductive tissue of transgenic plants, recombinant DNA molecules useful in such methods, and transgenic cells, seeds, and plants containing such recombinant DNA molecules. The invention provides an improvement on the art by providing male tissue-specific siRNA target elements (mts-siRNAs) capable of providing enhanced selective regulation of the expression of a protein encoded by a transcribable polynucleotide molecule, and provides recombinant DNA molecules and compositions comprising such mts-siRNA target elements, and methods for using such mts-siRNA target elements to induce male sterility in transgenic plants for the production of hybrid seeds. In one aspect, the invention provides a recombinant DNA molecule comprising an mts-pRNA target element operatively linked to a heterologous transcribable polynucleotide molecule. In one embodiment, the mts-pRNA target element is included within a 3' untranslated region operatively linked to the heterologous transcribable polynucleotide molecule. In another embodiment, the mts-pRNA target element is located between the heterologous transcribable polynucleotide molecule and an operatively linked polyadenylation sequence that forms part of a 3' untranslated region. In one embodiment, the mts-pRNA target element comprises a sequence selected from the group consisting of SEQ ID NO: 1-16, 23-92 and complements thereof. In another embodiment, the heterologous transcribable polynucleotide molecule confers herbicide tolerance, for example, RQonnn / i ζπζ / β / υιλι vegetative herbicide tolerance to a plant. In another embodiment, the heterologous transcribable polynucleotide molecule does not confer male reproductive herbicide tolerance to a plant. In another embodiment, the heterologous transcribable polynucleotide molecule is a glyphosate-tolerant 5-enolipyruvyl shikimate 3-phosphate synthase (EPSPS). In another aspect, the invention provides a recombinant DNA construct comprising an mts-ipRNA target element of the invention operatively linked to a heterologous transcribable polynucleotide molecule. In another aspect, the invention provides a method for producing a recombinant DNA molecule comprising operatively attaching at least one mts-pRNA target element to a heterologous transcribable polynucleotide molecule. In one embodiment, the mts-pRNA target element comprises a sequence selected from the group consisting of SEQ ID NO: 1-16, 23-92 and complements thereof. In another aspect, the invention provides a transgenic plant comprising an mts-pRNA target element of the invention. In one embodiment, the transgenic plant comprises the mts-pRNA target element operatively linked to a heterologous transcribable polynucleotide molecule. In another embodiment, the mts-pRNA target element comprises a sequence selected from the group consisting of SEQ ID NO: 1-16, 23-92 and complements thereof. In another embodiment, the transgenic plant is produced by transforming a plant with a recombinant DNA molecule or a DNA construct comprising at least one mts-pRNA target element operatively linked to a heterologous transcribable polynucleotide molecule. In another aspect, the invention provides a seed, cell, or part of said transgenic plant. In one embodiment, the plant is a monocotyledonous plant. In another embodiment, the plant is a maize plant (Zea mays). In another aspect, the invention also provides a method for selectively regulating the expression of a protein in the male reproductive tissue of a transgenic plant by expressing in the transgenic plant a recombinant DNA molecule comprising an mts-siRNA target element operatively linked to a heterologous transcribable polynucleotide molecule. In one embodiment, the mts-siRNA target element comprises a sequence selected from the group consisting of SEQ ID NO: 1-16, 23-92 and complements thereof. In another embodiment, the heterologous transcribable polynucleotide molecule confers herbicide tolerance, e.g., vegetative herbicide tolerance, to a plant. In another embodiment, the heterologous transcribable polynucleotide molecule does not confer herbicide tolerance to the male reproductive tissue of a plant.In another modality, the polynucleotide molecule that can be heterologously transcribed is a glyphosate-tolerant EPSPS. In another aspect, the invention provides a method for inducing sterility RQonnn / i ζπζ / ε / υιλι male in a transgenic plant, including the step of applying herbicide to a transgenic plant having in its genome a recombinant DNA molecule comprising an mts-ipRNA target element operatively linked to a heterologous transcribable polynucleotide molecule that confers tolerance to at least one first herbicide to the transgenic plant, wherein the herbicide is applied before or concurrently with the development of the male reproductive tissue of the transgenic plant, whereby male sterility is induced in the transgenic plant. In one embodiment, the heterologous transcribable polynucleotide molecule confers tolerance to vegetative herbicides, but does not confer tolerance to male reproductive herbicides to the transgenic plant. In another embodiment, the transgenic plant is a maize plant.In another embodiment, the application of the herbicide prevents at least pollen development, pollen dispersal, or anther extrusion in the treated transgenic plant. In another embodiment, the stage of male reproductive tissue development during which the herbicide is applied is a selected stage from the group consisting of stages V4, V5, V6, V7, V8, V9, V10, V11, V12, V13, and V14 of maize plant development. In another modality, the herbicide is selected from the group consisting of acetyl coenzyme A carboxylase (ACCase) inhibitors, acetolactate synthase (ALS) inhibitors, photosystem II (PSII) inhibitors, protoporphyrinogen oxidase (PPO) inhibitors, 4-hydroxyphenyldioxygenase (HPPD) inhibitors, 5-enolipyruvyl shikimate 3-phosphate synthase (EPSPS) inhibitors, glutamine synthetase (GS) inhibitors, and synthetic auxins.In another modality, the herbicide is glyphosate and the polynucleotide that can be transcribed heterologous encodes a glyphosate-tolerant EPSPS. In one aspect, the invention also provides a method for producing a hybrid seed comprising applying an effective amount of a herbicide to a transgenic plant comprising in its genome a recombinant DNA molecule comprising an mts-ipRNA target element operatively linked to a heterologous transcribable polynucleotide molecule, wherein the herbicide is applied before or concurrently with the development of the male reproductive tissue of the transgenic plant, thereby inducing male sterility in the transgenic plant; fertilizing the transgenic plant with pollen from a second plant; and allowing the hybrid seed to form from the transgenic plant. In one embodiment, the transgenic plant is maize. In another embodiment, the herbicide is glyphosate and the heterologous transcribable polynucleotide molecule is a glyphosate-tolerant EPSPS.In one embodiment, glyphosate is applied concurrently with development at an effective rate of approximately 0.125 pounds of acid equivalent per acre and approximately 8 pounds of acid equivalent per acre. In another method, the invention provides a hybrid seed produced by said method. In one embodiment, the hybrid seed comprises the recombinant DNA molecule. Other specific embodiments of the invention are described in the following RQonnn / i ζπζ / ε / υιλι detailed description. Throughout this descriptive memorandum and the claims, unless the context requires otherwise, the word understand and its variations such as comprises or comprising implies the inclusion of an integer, element or stage, or group of integers, elements or stages, but not the exclusion of any other integer, element or stage, or group of integers, elements or stages. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1. Illustration of the tassel development stages V7 in T2, V8 / V9 in T4, V10 / V11 in T5, V12 in T6, and VT in T7, showing tassel size and morphology, with photographs of the lower panels of anther cross-sections showing the pollen development stage. The stages previously used (V10 / V11 and V12) in the technique for isolating small RNA molecules for the identification of mts-siRNA molecules are indicated by a solid-lined box; the stages (V7, V8 / V9, V10 / V11, and V12) described herein for isolating small RNA molecules are indicated by a dashed-lined box. Figure 2. Graphical representation of mts-pRNA sequence alignment on the cDNA sequence provided as SEQ ID NO: 17. The cDNA sequence is indicated from nucleotide 1 to 1826, and the short lines represent the alignment of the complementary strand of individual mts-pRNA sequences, adjacent mts-pRNA sequence segments, or overlapping mts-pRNA sequences (relatively longer lines) to the cDNA sequence. mts-pRNA sequences that have the same strand as cDNA are not shown. The normalized relative expression level of mts-pRNA is indicated on the left. The area within the box represents a region of the cDNA sequence rich in mts-pRNA targets. Figure 3. Diagram illustrating double-stranded mts-pRNA, single-stranded mts-pRNA, mts-pRNA target sequence within an mRNA, and an mRNA region with a high number of mts-pRNA target sequences useful as an mts-pRNA target element. Figure 4. Photograph of R0 maize tassel and Alexandar ink-stained pollen from double-copy transgenic plants containing a recombinant DNA molecule comprising a transgene encoding a glyphosate-tolerant EPSPS protein operatively linked to an mts-ipRNA target element (SEQ ID NO: 2). Events 1–4 were sprayed with 0.75 lb ae / acre of glyphosate herbicide at V5 followed by V8 and showed tassels with complete male sterility as defined by the absence of anthesis and non-viable or no pollen grains detected in the anthers. Control plants did not receive a glyphosate application and showed normal anthesis and pollen dispersal. RQonnn / i zciz / r / yiai microscopic observation detected normal pollen grains. BRIEF DESCRIPTION OF THE SEQUENCES SEQ ID NO: 1 - An mts-siRNA target element sequence having 95% sequence identity with nucleotide positions 1429 to 1628 of the cDNA sequence provided herein as SEQ ID NO: 17. SEQ ID NO: 2 - An mts-siRNA target element sequence having 95% sequence identity with nucleotide positions 1429 to 1628 of the cDNA sequence provided herein as SEQ ID NO: 17 and having a single nucleotide change (T69A) with respect to SEQ ID NO: 1. SEQ ID NO: 3 - An mts-siRNA target element sequence corresponding to nucleotide positions 239 to 433 of the cDNA sequence provided herein as SEQ ID NO: 18. SEQ ID NO: 4 - An mts-siRNA target element sequence corresponding to nucleotide positions 477 to 697 of the cDNA sequence provided herein as SEQ ID NO: 18. SEQ ID NO: 5 - An mts-siRNA target element sequence corresponding to nucleotide positions 239 to 433 of the cDNA sequence provided herein as SEQ ID NO: 19. SEQ ID NO: 6 - An mts-siRNA target element sequence corresponding to nucleotide positions 370 to 477 of the cDNA sequence provided herein as SEQ ID NO: 19. SEQ ID NO: 7 - An mts-siRNA target element sequence corresponding to nucleotide positions 1357 to 1562 of the cDNA sequence provided herein as SEQ ID NO: 20. SEQ ID NO: 8 - An mts-siRNA target element sequence corresponding to nucleotide positions 247 to 441 of the cDNA sequence provided herein as SEQ ID NO: 21. SEQ ID NO: 9 - The reverse complement of an mts-ipRNA target element sequence having 99% sequence identity with nucleotide positions 191 to 490 of the cDNA sequence provided herein as SEQ ID NO: 22 with three nucleotide mismatch errors (C314A, A350G and G408A). SEQ ID NO: 10-16 - Recombinant mts-ipRNA target element sequences. SEQ ID NO: 17 - cDNA sequence containing at least one region rich in AQQnnn / ι ζπζ / ε / υιλι mts-siRNA target sequence. It contains the mts-siRNA target element sequences represented by SEQ ID NO: 1 and SEQ ID NO: 2 and aligns with the mts-siRNA sequences SEQ ID NO: 23-31. SEQ ID NO: 18 - cDNA sequence containing at least one mts-pRNA target sequence rich region. Contains the mts-pRNA target element sequences represented by SEQ ID NO: 3 and SEQ ID NO: 4 and aligns with the mts-pRNA sequences SEQ ID NO: 32-42. SEQ ID NO: 19 - cDNA sequence containing at least one mts-pRNA target sequence rich region. Contains the mts-pRNA target element sequences represented by SEQ ID NO: 5 and SEQ ID NO: 6 and aligns with the mts-pRNA sequences SEQ ID NO: 43-52. SEQ ID NO: 20 - cDNA sequence containing at least one mts-pRNA target sequence-rich region. Contains the mts-pRNA target element sequence represented by SEQ ID NO: 7 and aligns with mts-pRNA sequences SEQ ID NO: 53-60. SEQ ID NO: 21 - cDNA sequence containing at least one mts-pRNA target sequence-rich region. Contains the mts-pRNA target element sequence represented by SEQ ID NO: 8 and aligns with mts-pRNA sequences SEQ ID NO: 61-69. SEQ ID NO: 22 - cDNA sequence containing at least one mts-pRNA target sequence-rich region. Contains the mts-pRNA target element sequence represented by SEQ ID NO: 9 and aligns with mts-pRNA sequences SEQ ID NO: 70-87. SEQ ID NO: 23-92 - DNA sequences corresponding to the mtsRNAp sequences of the invention, which align with the cDNA sequences provided herein as SEQ ID NO: 17-22. DETAILED DESCRIPTION OF THE INVENTION The invention provides recombinant DNA molecules, compositions, and methods for selectively regulating protein expression, for example, the expression of a heterologous transtranscribable polynucleotide molecule, in the male reproductive tissue of a transgenic plant, and their uses. In one aspect, the invention provides a recombinant DNA molecule that includes a male tissue-specific small interfering RNA (mts-pRNA) target element operatively linked to a heterologous transtranscribable polynucleotide. Such recombinant DNA molecules are useful for selectively regulating the expression of a heterologous transtranscribable polynucleotide in the male reproductive tissue of a transgenic plant. The nucleic acid sequences can be provided as either DNA or RNA. RQQnnn / ι ζπζ / ε / υιλι as specified; the description of one necessarily defines the other, as known by a person skilled in the art. Furthermore, the description of a given nucleic acid sequence necessarily defines and includes the complement of that sequence, as known by a person skilled in the art. Small interfering RNA (siRNA) is a class of RNA molecules approximately 18–26 nucleotides (nt) in length (e.g., 18, 19, 20, 21, 22, 23, 24, 25, or 26 nt). An siRNA sequence can be represented using the RNA nucleotide sequence consisting of guanine (G), cytosine (C), adenine (A), and uracil (U) or using the equivalent DNA nucleotide sequence of guanine (G), cytosine (C), adenine (A), and thymine (T). siRNA functions within RNA-induced silencing complexes (RISCs) to trigger sequence-specific degradation of messenger RNA (mRNA), resulting in altered gene expression and downregulation of the gene-encoded protein. A male tissue-specific siRNA, or mts-siRNA, is a siRNA enriched or expressed specifically in the male reproductive tissue (e.g., the male inflorescence) of a plant and therefore has a male tissue-specific expression pattern. Male tissue-specific siRNAs have been identified in plants and can be detected using techniques known in the art, such as low molecular weight Northern blotting. An mts-siRNA sequence is the nucleic acid sequence of an mts-siRNA. Exemplary mts-siRNA sequences in the form of the corresponding DNA sequence of the double-stranded mts-siRNA molecule are provided herein as SEQ ID NO: 23-92. A DNA sequence that is complementary to an mts-npRNA sequence is referred to herein as an mts-npRNA target. The mts-npRNA target is contained within the DNA sequence of a gene and is transcribed into the RNA sequence of the corresponding mRNA molecule. A single strand of a double-stranded mts-npRNA molecule can bind, or hybridize, under typical physiological conditions to the mts-npRNA target in the mRNA molecule. See Figure 3. A nucleic acid sequence is complementary to an mts-npRNA sequence if alignment of the two nucleic acid sequences yields an exact match (no mismatches, i.e., complete complement), one mismatch, two mismatches, or three mismatches along the length of the mts-npRNA sequence.Complementary sequences can have base pairing with each other according to standard Watson-Crick complementarity rules (i.e., guanine with cytosine (G:C) pairs and adenine with either thymine (A:T) or uracil (A:U) pairs). An mts-pRNA target sequence is the nucleic acid sequence of an mts-pRNA target. Exemplary mts-pRNA target sequences are provided herein as SEQ ID NO: 23-92. More than one mts-pRNA target can be clustered or even overlapped within a single DNA molecule. A DNA molecule comprising more than one mts-pRNA target RQonnn / i ζπζ / ε / υιλι is referred to herein as an mts-pRNA target element. An mts-pRNA target element comprises at least two or more mts-pRNA targets within a 500-nucleotide sequence window. An mts-pRNA target element can be of any length, for example, approximately 30 nucleotides (nt), approximately 40 nt, approximately 50 nt, approximately 60 nt, approximately 70 nt, approximately 80 nt, approximately 90 nt, approximately 100 nt, approximately 150 nt, approximately 200 nt, approximately 250 nt, approximately 300 nt, approximately 350 nt, approximately 400 nt, approximately 450 nt, or approximately 500 nt. An mts-ipRNA target element sequence is the nucleic acid sequence of an mts-ipRNA target element. Exemplary mts-ipRNA target element sequences are provided herein as SEQ ID NO: 1-16. As used herein, a recombinant DNA molecule, polypeptide, protein, cell, or organism may be non-natural or a human-made creation using the tools of genetic engineering and, as such, is the product of human activity and is not normally found in nature. A recombinant DNA molecule refers to a DNA molecule comprising a combination of DNA sequences or molecules that are not found together in nature without human intervention. For example, a recombinant DNA molecule may be a DNA molecule composed of at least two heterologous DNA molecules, a DNA molecule comprising a DNA sequence that deviates from DNA sequences existing in nature, or a DNA molecule that has been incorporated into the DNA of a host cell through genetic transformation.In one embodiment, a recombinant DNA molecule of the invention is a DNA molecule comprising an mts-siRNA target element operatively linked to at least one transcribable polynucleotide molecule, for example, wherein the transcribable polynucleotide molecule is heterologous to the mts-siRNA target element. As used herein, a recombinant molecule, cell, or organism may be synthetic. As used herein, the term heterologous refers to the combination of two or more DNA or protein molecules when such a combination is not normally found in nature or when such a combination is provided in an orientation or order that differs from that found in nature. For example, the two DNA molecules may be derived from different species or created synthetically, and / or the two DNA molecules may be derived from different genes, e.g., different genes from the same species or the same genes from different species. In one example, a regulatory element or mts-ipRNA target element may be heterologous with respect to an operationally bound, transcribable polynucleotide molecule if such a combination is not normally found in nature; that is, the transcribable polynucleotide molecule is not found in nature bound together. RQQnnn / ι ζπζ / β / υιλι operatively to the regulatory element or the mts-ipRNA target element. In one embodiment, such heterologous combination may comprise an mts-ipRNA target element that may be plant-derived or chemically synthesized and may be operatively linked to a transcribable polynucleotide molecule, such as a bacterial transgene encoding a protein for herbicide tolerance, such as cp4-EPSPS (e.g., as provided herein as SEQ ID NO: 93). Furthermore, a particular sequence may be heterologous with respect to a cell or organism into which it is introduced (e.g., a sequence not naturally found in that particular cell or organism). As used herein, the term "isolated" means separated from other molecules that are typically associated with it in its natural state. For example, an isolated DNA molecule is a molecule that is present alone or combined with other compositions, but is not in its natural genomic location or state. In one sense, the term "isolated" refers to a DNA molecule that is separated from the nucleic acids that normally flank the DNA molecule in its natural state. For example, an isolated DNA molecule may be a DNA molecule composed of at least two heterologous DNA molecules. In another example, an isolated DNA molecule may be a DNA molecule that has been incorporated into a novel genomic location in a host cell by genetic transformation.Therefore, a DNA molecule fused or operationally joined to one or more different DNA molecules with which it would not be found associated in nature—for example, as a result of plant transformation techniques or recombinant DNA—is considered isolated herein. These molecules are considered isolated even when they are integrated into the chromosome of a host cell or present in a nucleic acid solution with other DNA molecules. The term operatively bound refers to at least two nucleotide molecules arranged or linked in such a way that one can affect the function of the other. The two nucleotide molecules may be part of a single contiguous nucleotide molecule and may be adjacent or separate. For example, a mts-siRNA target element may be operatively bound to a transcribable polynucleotide molecule. In one modality, an operatively bound mts-siRNA molecule can affect the transcription, translation, or expression of the transcribable polynucleotide molecule.For example, an mts-pRNA target element is operatively bound to a transcribable polynucleotide molecule. If, after transcription in a male reproductive tissue cell, the presence of the mts-pRNA target element on the mRNA molecule triggers the regulation of the expression of the transcribable polynucleotide molecule in the cell, induced by endogenous mts-pRNA and the RISC pathway, the operative binding of the mts-pRNA target element and the molecule is described. The incorporation of a transcribable mts-pRNA target element can be achieved, for example, by incorporating an mts-pRNA target element adjacent to the transcribable polynucleotide molecule (e.g., located 5' or 3' with respect to the transcribable polynucleotide molecule, but not necessarily contiguously attached), or adjacent to an untranslated region (UTR) of the polynucleotide molecule (e.g., located in or next to the 5' UTR or the 3' UTR), and / or 3' with respect to the transcribable polynucleotide molecule and 5' with respect to the polyadenylation signal. In one modality, an mts-pRNA target element is located between the transcribable polynucleotide molecule and the polyadenylation sequence, i.e., 3' and adjacent to the transcribable polynucleotide molecule.In another conformation, an mts-pRNA target element is located between the stop codon of the transcribable polynucleotide molecule and the polyadenylation sequence. In yet another conformation, an mts-pRNA target element is located within the 3' UTR sequence adjacent to the transcribable polynucleotide molecule. This paper provides examples of the identification of mts-pRNAs, mts-pRNA targets, and mts-pRNA target elements. These can be identified using methods known to those skilled in the art, such as bioinformatics analysis of plant RNA and cDNA libraries. In particular, mts-pRNAs can be identified from RNA libraries and sequenced. The identified mts-pRNA sequences can then be compared with cDNA and / or genomic sequence collections to identify mts-pRNA targets and mts-pRNA target elements useful for the development of recombinant DNA molecules and constructs, as described herein. In some embodiments, mts-pRNA target elements are created, synthesized, or modified in vitro. For example, mts-pRNA target elements can be modified to contain more, fewer, or different mts-pRNA target sequences, or to rearrange the relative position of one or more mts-pRNA target sequences. In some embodiments, such modification can be beneficial in increasing or decreasing the effect of the mts-pRNA target element. The methods for creating, synthesizing, or modifying an mts-pRNA target element in vitro, and for determining the optimal variation for the desired level of regulation, are known to those skilled in the art.Exemplary mts-pRNA target elements may be created by combining DNA sequences, or fragments thereof, from two or more mts-pRNA targets, two or more mts-pRNA target elements, two or more mts-pRNA target-rich cDNA regions, or one or more mts-pRNA targets or fragments of two or more mts-pRNA target-rich cDNA regions, for example by combining all or fragments of two or more mts-pRNA target elements provided herein as SEQ ID NO: 1-9, or by combining two or more of the mts-pRNA sequences provided herein as. RQonnn / i ζπζ / ε / υιλι SEQ ID NO: 23-92, or by combining all or fragments of two or more mts-ipRNA target elements provided herein as SEQ ID NO: 1-9 with one or more of the mts-ipRNA sequences provided herein as SEQ ID NO: 23-92. Such exemplary recombinant mts-ipRNA target elements are provided herein as SEQ ID NO: 10-16. The DNA sequence of the mts-pRNA target element can also be varied by incorporating 1-3 nucleotide mismatches into an mts-pRNA target sequence (with respect to a given mts-pRNA sequence). In another embodiment, the present invention includes recombinant DNA molecules or mts-pRNA target elements having at least approximately 80% sequence identity, approximately 85% sequence identity, approximately 90% sequence identity, approximately 91% sequence identity, approximately 92% sequence identity, approximately 93% sequence identity, approximately 94% sequence identity, approximately 95% sequence identity, approximately 96% sequence identity, approximately 97% sequence identity,approximately 98% sequence identity and approximately 99% sequence identity with any of the DNA molecules, mts-ipRNA target elements (e.g. SEQ ID NO: 1-9), recombinant mts-ipRNA target elements (e.g. SEQ ID NO: 10-16) or cDNA sequences (e.g. SEQ ID NO: 17-22) of the present invention. In another embodiment, the present invention provides fragments of a DNA molecule described herein. Such fragments may be useful as mts-ipRNA target elements or may be combined with other mts-ipRNA target elements, mts-ipRNA sequences, or fragments thereof for the construction of recombinant mts-ipRNA target elements, as described above. In specific embodiments, such fragments may comprise at least approximately 20, at least approximately 30, at least approximately 40, at least approximately 50, at least approximately 60, at least approximately 70, at least approximately 80, at least approximately 90, at least approximately 100, at least approximately 110, at least approximately 120, at least approximately 130, at least approximately 140, at least approximately 150, at least approximately 160, at least approximately 170, at least approximately 180, at least approximately 190,at least approximately 200, at least approximately 210, at least approximately 220, at least approximately 230, at least approximately 240, at least approximately 250, at least approximately 260, at least approximately 270, at least approximately 280, at least approximately 290, at least approximately 300, at least approximately 350, at least approximately 400, at least approximately 450, at least approximately 500 contiguous, or longer, nucleotides of a DNA molecule described herein, for example, RQQnnn / ι ζπζ / β / υιλι is a target element of mts-siRNA or cDNA sequence described herein. The methods for producing such fragments from an initial DNA molecule are known in the art. The effectiveness of the modifications, duplications, deletions, or rearrangements described herein on the desired expression aspects of a particular transcribable polynucleotide molecule can be empirically evaluated in stable and transient plant assays, such as those described in the working examples herein, to validate the results, which may vary depending on the changes made and the objective of the change in the initial DNA molecule. An mts-pRNA target and an mts-pRNA target element can function in any direction, meaning it is non-directional, and as such can be used in either the 5' to 3' or 3' to 5' orientation in a recombinant DNA molecule or DNA construct. As used herein, expression of a transcribable polynucleotide molecule or expression of a protein refers to the production of a protein from a transcribable polynucleotide molecule and the resulting transcript (mRNA) in a cell. The term protein expression, therefore, refers to any pattern or translation of a transcribed RNA molecule into a protein molecule. Protein expression may be characterized by its temporal, spatial, developmental, or morphological qualities, as well as by quantitative or qualitative indications. In one embodiment, the recombinant DNA molecule of the invention may be used to selectively regulate the expression of a protein or transcribable polynucleotide molecule in male reproductive tissues of a transgenic plant.In this modality, the expression of the recombinant DNA molecule in a transgenic plant can lead to the expression of an operationally linked polynucleotide molecule in at least vegetative tissues, but not in male reproductive tissues. In certain modalities, this regulation of protein expression refers to suppression or reduction; for example, suppressing or reducing the level of protein produced in a cell, for example, through RNAi-mediated post-translational genetic regulation. Selective regulation of protein expression, as used herein, refers to a reduction in protein production in a cell or tissue compared to a reference cell or tissue of at least approximately 75%, at least approximately 80%, at least approximately 85%, at least approximately 90%, at least approximately 95%, at least approximately 99%, or 100% reduction (i.e., a complete reduction). A reference cell or tissue may be, for example, a plant cell or tissue from the same or a similar transgenic plant that expresses the protein, or a cell or tissue from a transgenic plant that has a similar transgene encoding the protein but lacks the specific transgene that encodes the protein. RQonnn / i zciz / r / yiai of an operationally bound mts-ipRNA target element. The regulation of protein expression can be determined using any technique known to the person skilled in the art, for example, by direct measurement of protein accumulation in a cell or tissue sample using a technique such as ELISA or Western blot analysis, by measuring the protein's enzymatic activity, or by phenotypic determination of protein expression. In one modality, selective regulation of protein expression refers to a sufficient reduction in the expression of a protein capable of conferring herbicide tolerance in the male tissue of a transgenic plant to produce a detectable phenotype of altered male fertility in a transgenic plant to which herbicide is applied as an induced sterility spray.The detection of altered male fertility in this transgenic plant, therefore, indicates the selective regulation of protein expression. As used herein, the term transgene encoding a recombinant protein or transcribable polynucleotide molecule refers to any nucleotide molecule capable of being transcribed into an RNA molecule, including, but not limited to, those having a nucleotide sequence that encodes a polypeptide sequence. Depending on the conditions, the nucleotide sequence may or may not be translated into a polypeptide molecule in a cell. The boundaries of a transgene or transcribable polynucleotide molecule are commonly defined by a translation start codon at the 5' end and a translation stop codon at the 3' end. The term transgene refers to a DNA molecule artificially incorporated into the genome of a host organism or cell, in the current or any previous generation of the organism or cell, as a result of human intervention, for example, through plant transformation methods. As used herein, the term transgenic means comprising a transgene; for example, a transgenic plant refers to a plant that comprises a transgene in its genome, and a transgenic trait refers to a characteristic or phenotype carried or conferred by the presence of a transgene incorporated into the plant's genome. As a result of such genomic alteration, the transgenic plant is distinctly different from the related wild-type plant, and the transgenic trait is a trait not found naturally in the wild-type plant.The transgenic plants of the invention comprise the recombinant DNA molecule provided by the invention. A transgene or transcribable polynucleotide molecule of the invention includes, among others, a transgene or transcribable polynucleotide molecule that provides a desirable characteristic associated with the morphology, physiology, growth, development, yield, nutritional properties, disease resistance, pest resistance, herbicide tolerance, stress tolerance, environmental stress tolerance, or chemical tolerance of the RQonnn / i ζπζ / β / υιλι plant. In one embodiment, a transcribable polynucleotide molecule of the invention encodes a protein that, when expressed in a transgenic plant, confers herbicide tolerance in at least one cell and / or tissue where the expressed protein is produced; selective regulation of the herbicide tolerance protein in the male reproductive tissue of the transgenic plant together with timely herbicide application results in at least induced reduced male fertility or induced male sterility. Such inducible male sterility combined with tolerance to vegetative herbicides can be used to increase the efficiency with which hybrid seed is produced, for example, by eliminating or reducing the need to physically castrate the maize plant used as a female plant in a given cross during hybrid seed production. Herbicide-inducible male sterility systems have been described, for example, in U.S. Patent No. 6,762,344; U.S. Patent No. 8,618,358; and U.S. Patent Publication 2013 / 0007908.Some examples of herbicides useful for the practice of the invention include, among others, acetyl coenzyme A carboxylase (ACCase) inhibitors (e.g., fops and dims), acetolactate synthase (ALS) inhibitors (e.g., sulfonylureas (SU) and imidazolinones (IMI)), photosystem II (PSII) inhibitors (e.g., trayazines and phenyl ethers), protoporphyrinogen oxidase (PPO) inhibitors (e.g., flumioxazin and fomesafen), 4-hydroxyphenyl pyruvate dioxygenase (HPPD) inhibitors (e.g., isoxaflutol and triketones such as mesotrione), 5-enolipyruvyl shikimate 3-phosphate synthase (EPSPS) inhibitors (e.g., glyphosate), glutamine synthetase (GS) inhibitors (e.g., glufosinate and phosphinothricin), synthetic auxins (e.g., 2,4-D and dicamba).Some examples of transgenes or polynucleotide molecules that can be transcribed for use in the practice of the invention include, among others, genes encoding proteins that confer tolerance to HPPD inhibitors (e.g., herbicide-sensitive HPPD), genes encoding proteins that confer tolerance to glufosinate (e.g., pat and bar), genes encoding proteins that confer tolerance to glyphosate (e.g., a glyphosate-tolerant EPSPS, such as cp4-epsps, provided herein as SEQ ID NO: 93), and genes encoding proteins that confer tolerance to a synthetic auxin such as dicamba (e.g., dicamba monooxygenase (DMO)) and 2,4-D (e.g., gene ( / ?>dichlorprop dioxygenase (rdpA)). The recombinant DNA constructs of the invention may include the recombinant DNA molecules of the invention and are made using techniques known in the art and, in various embodiments, are incorporated into plant transformation vectors, plasmids, or plastid DNA. Such recombinant DNA constructs are useful for producing transgenic plants and / or cells and, as such, may also be contained within the genomic DNA of a transgenic plant, seed, cell, or plant part. Therefore, the present invention includes embodiments RQonnn / i zctz / r / yiai wherein the recombinant DNA construct is located within a plant transformation vector, in a biolistic particle for transforming a plant cell, within a chromosome or plastid of a transgenic plant cell, within a transgenic cell, transgenic plant tissue, transgenic plant seed, transgenic pollen grain, or a transgenic or partially transgenic plant (e.g., with grafting). A vector is any DNA molecule that can be used for the purpose of transforming the plant, i.e., introducing DNA into a cell. The recombinant DNA constructs of the invention, for example, can be inserted into a plant transformation vector and used for plant transformation to produce transgenic plants, seeds, and cells. Methods for constructing plant transformation vectors are known in the art.The plant transformation vectors of the invention generally include, among others: a promoter suitable for the expression of an operationally bound DNA, an operationally bound recombinant DNA construct, and a polyadenylation signal (which may be included in a 3' UTR sequence). Promoters useful for the practice of the invention include those that function in a plant for the expression of an operationally bound polynucleotide. Such promoters are varied and known in the art and include those that are inducible, viral, synthetic, constitutive, time-regulated, spatially regulated, and / or spatio-temporally regulated. Some additional optional components include, among others, one or more of the following targets: a 5' UTR, an enhancer, a cis-acting target, an intron, a signal sequence, a transit peptide sequence, and one or more selectable marker genes.In one embodiment, a plant transformation vector comprises a recombinant DNA construct. The recombinant DNA constructs and plant transformation vectors of the present invention are made by any method suitable for the intended application, taking into account, for example, the type of expression desired, the transgene or polynucleotide molecule that can be transcribed, and the suitability of its use in the plant in which the recombinant DNA construct is expressed. General methods useful for manipulating DNA molecules to make and use recombinant DNA constructs and plant transformation vectors are known in the art and are described in detail, for example, in laboratory guides and manuals including Michael R. Green and Joseph Sambrook, Molecular Cloning: A Laboratory Manual (fourth edition) ISBN: 978-1-936113-42-2, Coid Spring Harbor Laboratory Press, NY (2012). The recombinant DNA molecules and constructs of the invention can be modified by methods known in the art, either wholly or in part, for example, to increase the convenience of DNA manipulation (such as restricting enzyme recognition sites or recombination-based cloning sites), or to include RQonnn / i zciz / r / yiai preferred plant sequences (such as plant codon usage or Kozak consensus sequences) or to include sequences useful for the design of recombinant DNA molecules and constructs (such as spade or linker sequences). In certain embodiments, the DNA sequence of the molecule and recombinant DNA construct includes a DNA sequence that has been codon-optimized for the plant in which the molecule or recombinant DNA construct is expressed. For example, a recombinant DNA molecule or construct expressed in a plant may have its entire sequence or parts of it codon-optimized for expression in that plant using methods known in the art.The recombinant DNA molecules or constructs of the invention can be stacked with other recombinant DNA molecules or transgenic events to impart additional traits (e.g., in the case of transformed plants, traits include herbicide resistance, pest resistance, cold germination tolerance, water deficit tolerance) by, for example, expressing or regulating other genes. One aspect of the invention includes transgenic plant cells, transgenic plant tissues, and transgenic seeds or plants containing a recombinant DNA molecule of the invention. A further aspect of the invention includes artificial or recombinant plant chromosomes containing a recombinant DNA molecule of the invention. Suitable methods for transforming host plant cells for use with the present invention include virtually any method by which DNA can be introduced into a cell (e.g., when a recombinant DNA molecule is stably integrated into a plant chromosome) and are known in the art. An exemplary and widely used method for introducing a recombinant DNA molecule into plants is the Agrobacterium transformation system, which is known to those skilled in the art.Another exemplary method for introducing a recombinant DNA molecule into plants is the insertion of a recombinant DNA molecule into a plant genome at a predetermined site using site-directed integration methods. Site-directed integration can be achieved through any known method in the art, for example, by using zinc finger nucleases, genetically modified or naturally occurring meganucleases, TALE-endonucleases, or an RNA-guided endonuclease (e.g., a CRISPR / Cas9 system). Transgenic plants can be regenerated from a transformed plant cell using plant cell culture methods. A transgenic plant homozygous for a transgene can be obtained by sexual mating (self-fertilization) of an independently segregating transgenic plant containing a single self-aggregated gene sequence, for example, an R0 or F0 plant, to produce R1 or F1 seed.One-quarter of the R1 or F1 seed produced is homozygous for the transgene. Plants grown from germinated R1 or F1 seed can be tested for heterozygosity, typically using an SNP assay. RQQnnn / i ζπζ / ε / υιλι a thermal amplification test that allows the distinction between heterozygotes and homozygotes (i.e., a zygosity test). The invention provides a transgenic plant having in its genome a recombinant DNA molecule of the invention, including, without limitation, alfalfa, cotton, corn, sorghum, rice, soybean, and wheat, among others. The invention also provides plant cells, plant parts, and progeny of such a transgenic plant. As used herein, progeny includes any plant, seed, plant cell, and / or plant part produced or regenerated from a plant, seed, plant cell, and / or plant part that includes a recombinant DNA molecule of the invention. Transgenic plants, cells, parts, progeny plants, and seeds produced from such plants may be homozygous or heterozygous for the recombinant DNA molecule of the invention. Plant parts of the present invention may include, among others, leaves, stems, roots, seeds, endosperm, ovule, and pollen.The parts of a plant of the invention may be viable, non-viable, regenerable, or non-regenerable. The invention also includes and provides transformed plant cells comprising a DNA molecule of the invention. The transformed or transgenic plant cells of the invention include both regenerable and non-regenerable plant cells. The present invention further includes embodiments in which the recombinant DNA molecule is contained in a commodity produced from a transgenic plant, seed, or plant part of the present invention; such commodity products include, among others, harvested parts of a plant, ground or whole grains or seeds of a plant, or any food or non-food product comprising the recombinant DNA molecule of the present invention. The invention provides a method for inducing male sterility in a transgenic plant comprising (a) cultivating a transgenic plant comprising a recombinant DNA molecule comprising a heterologous transcribable polynucleotide molecule conferring herbicide tolerance operatively linked to an mts-ipRNA target element, and (b) applying an effective amount of the herbicide to the transgenic plant to induce male sterility. An effective amount of a herbicide is an amount sufficient to render a transgenic plant comprising a recombinant DNA molecule of the invention male-sterile. In one embodiment, an effective amount of glyphosate is between approximately 0.125 pounds of acid equivalent per acre and approximately 8 pounds of acid equivalent per acre.The herbicide application can be applied before or during the development of the male reproductive tissue, for example at a selected stage of the group consisting of the V4, V5, V6, V7, V8, V9, V10, V11, V12, V13 and V14 stages of corn plant development and can at least prevent pollen development, pollen dispersal and other extrusion. RQonnn / i ζπζ / ε / υιλι In one modality, the prevention of pollen development, pollen dispersal, and other extrusion can result from male sterility, and thus, the absence of pollen development, pollen dispersal, and other extrusion can be an indication of male sterility. However, in some cases, male-sterile plants may still produce small amounts of pollen. Therefore, in certain modalities, the presence of a small amount of pollen does not necessarily indicate male-fertile plants or the absence of male sterility. Plant development is usually determined on a stage scale based on the plant's growth. For maize, a common plant development scale used in the art is known as V-stages. V-stages are defined according to the uppermost leaf where the leaf collar is visible. VE corresponds to emergence, VI corresponds to the first leaf, V2 corresponds to the second leaf, V3 corresponds to the third leaf, and V(n) corresponds to the nth leaf. VT occurs when the last tassel branch is visible but before the silks emerge. When arranging a maize field, each specific V-stage is defined only when 50 percent or more of the plants in the field are at or beyond that stage. Other development scales are known to those skilled in the art and can be used with the methods of the invention. Another common tool for predicting and estimating corn growth and development stages is Growth Degree Units (GDUs). One factor in corn growth and development is heat. Heat is typically measured at a single point in time and expressed as temperature, but it can also be measured over a period and expressed as heat units. These heat units are commonly referred to as GDUs. GDUs can be defined as the difference between the average daily temperature and a selected base temperature subject to certain constraints. GDUs are calculated using the following equation: Growth Degree Unit = {(H + L) / 2}-B, where H is the daily high (but above 86°F), L is the daily low (but not below 50°F), and B is the base of 50°F.Because corn growth is slower when temperatures are above 86°F or below 50°F, limits are set on the daily high and daily low temperatures used in the formula. The lower cutoff for daily temperature also prevents the calculation of negative values. Therefore, if the daily high temperature exceeds 86°F, the daily low temperature used in the GDU formula is set to 86°F. Conversely, if the daily low temperature falls below 50°F, the daily low temperature used in the GDU formula is set to 50°F. If the daily high temperature does not exceed 50°F, no GDU is recorded for that day. The maximum GDU a corn plant can accumulate in a day is 36, and the minimum is zero. A corn plant's maturity grade is identified by the sum of its daily GDU values ​​over a specified time period. RQonnn / i zoz / β / uli Most corn seed producers use data from planting until physiological maturity, or the point at which grain filling is nearly complete. In most U.S. states, for example, cumulative GDUs are maintained for most geographic areas and are available from the USDA Crop Reporting Service or State Extension Services. Additionally, a tool for obtaining GDU information for a particular location is also described in U.S. Patent No. 6,967,656, which is incorporated herein by reference in its entirety. Another method for predicting the development of the clump for determining the timing of application of a male sterility-inducing herbicide is described in U.S. Patent No. 8,618,358, which is incorporated herein by reference in its entirety. The herbicides for use with the invention include any herbicide, including those active against acetyl coenzyme A carboxylase (ACCase), acetolactate synthase (ALS) inhibitors, photosystem II (PSII) inhibitors, protoporphyrinogen oxidase (PPO) inhibitors, 4-hydroxyphenyl pyruvate dioxygenase (HPPD) inhibitors, 5-enolipyruvyl shikimate 3-phosphate synthase (EPSPS) inhibitors, glutamine synthetase (GS) inhibitors, and synthetic auxins. The herbicides are known in the art and are described, for example, in Modern Crop Protection Compounds, Volume 1 (Second Edition), edited by Wolfgang Krámer, Ulrich Schirmer, Peter Jeschke, and Matthias Witschel, ISBN: 9783527329656, Wiley-VCH Verlag GmbH & Co. KGaA, Germany (2012). In one version, the herbicide is glyphosate. A hybrid seed can be produced using a method comprising (a) applying herbicide to a transgenic plant that includes a recombinant DNA molecule comprising a heterologous transcribable polynucleotide molecule conferring herbicide tolerance operatively linked to an mts-ipRNA target element, wherein the herbicide application is carried out during the development of the male reproductive tissue of the transgenic plant, thereby inducing male sterility in the transgenic plant; (b) fertilizing the transgenic plant with pollen from a second plant; and (c) harvesting hybrid seed from the transgenic plant. In one embodiment, the transgenic plant is maize. In another embodiment, the herbicide is glyphosate, and the protein encoded by the heterologous transcribable polynucleotide molecule is a glyphosate-tolerant EPSPS.In one modality, glyphosate is applied during the growing season at an effective rate of approximately 0.125 pounds of acid equivalent per acre and approximately 8 pounds of acid equivalent per acre. In another modality, the fertilization stage can be achieved by allowing natural fertilization, for example through wind pollination, or it can include mechanical or hand pollination. A hybrid seed can be harvested from a male-sterile transgenic plant that is RQonnn / i zciz / r / yiai has been pollinated from a second plant, wherein the male-sterile transgenic plant comprises a recombinant DNA molecule that includes a heterologous transcribable polynucleotide conferring herbicide tolerance operatively linked to an mts-ipRNA target element, and wherein the transgenic plant has been induced to be male-sterile by the application of an effective amount of herbicide during the development of male reproductive tissue. In one example, the herbicide is glyphosate and is applied during development at an effective amount of between approximately 0.125 pounds of acid equivalent per acre and approximately 8 pounds of acid equivalent per acre and prevents at least pollen development, pollen dispersal, or other extrusion. EXAMPLES The following examples describe improvements to hybrid seed production over that provided by the prior art. These improvements include novel mts-ipRNA targets and mts-ipRNA target elements for use in recombinant DNA molecules and transgenic plants to provide early-stage pollen development arrest, resulting in the absence of viable pollen grains in a wide range of germplasm, and related methods of use. The following examples are provided to demonstrate embodiments of the invention. EXAMPLE 1 identification of mts-ARNip targets Small RNA was isolated from four separate growth stages of tassel and three separate growth stages of ear of corn. See Table 1. Tassel-enriched small RNA was isolated from very early tassel developmental stages (V7, V8 / V9, V10 / V11, and V12) (see Figure 1). This yielded small RNA from younger male tissues than those previously used in the technique for obtaining tassel-enriched small RNA sequences. Small RNA libraries were prepared using the isolated small RNA, and high-throughput small RNA sequencing was performed on the libraries. Bioinformatics analysis was used to compare the sequences in these tassel and ear libraries with the sequences in small RNA libraries prepared from other maize tissues, including leaves collected at various growth stages, whole seedlings, roots collected at various growth stages, endosperm, and seed. This differential bioinformatics analysis identified thousands of tassel-enriched small RNA sequences with normalized expression in the range of RQonnn / i ζπζ / β / υιλι at 665 transcripts per quarter million (tpq). The identified tassel-enriched small RNA sequences are likely siRNAs due to their length (18–26 nucleotides) and their expected origin from a csRNA precursor. Due to male tissue specificity, these tassel-enriched small RNAs are referred to herein as male tissue-specific siRNAs (mts-siRNAs). RQonnn / i 7Π7 / β / υιλι TABLE 1 Description of small RNA libraries of tassel and corn cob Tassel / Ear Stage Tassel / Ear Size Tassel at microspore mother cell stage (plant at V7-V8) <1 cm Tassel at microspore mother cell stage-premeiosis (plant at V8-V9) 1-3 cm Tassel at early meiosis - microspore-free stage (plant at V9-V10) 3-17 cm Tassel at late stage - uninucleate microspores (plant at V12-VT) >17 cm Ear at premeiosis - four-nucleate immature embryo sac stage (plant at VT) 2-3 cm Ear at eight-nucleate immature embryo sac stage to later stage with up to 10 antipodal cells (plant at VT) 4-5 cm Ear at pollination stage (plant at VT-R1) 9-10 cm A real-time PCR method was used to identify and confirm that mts-pRNA sequences were specifically expressed in the tassel. Total RNA, including enriched small RNA, was extracted from the tissues listed in Table 1 and used to synthesize cDNA with reverse transcription primers consisting of 8 nt complementary to mts-pRNA sequences at the 3' end and a 35 nt universal sequence at the 5' end. After cDNA synthesis, real-time PCR was performed, where the sequence (14 to 18 nt) of one of the forward primers was identical to the 5' end of an mts-pRNA sequence, and the reverse primer was a universal primer. As an internal control, 18S RNA was amplified and used to normalize mts-pRNA levels. Real-time PCR data were used to limit the amount of mts-pRNA sequences that were enriched in the tassel. A microarray profiling assay of siRNA was performed using pParaflo® Microfluidics chips provided by LC Sciences LLC (Houston, Texas, USA) with 1,200 sequences selected from the thousands of mts-siRNA sequences identified from differential bioinformatics analysis. The microarray chips contained triplicate probes of the complementary sequence for each of the 1,200 mts-siRNA sequences. Total RNA was purified from 26 maize tissue pools (duplicate or triplicate tissue pools) from inbred plants LH244 (ATCC deposit number PTA-1173) or 01DKD2 (1294213) (ATCC deposit number PTA-7859). See Table 2. Each of the 26 RNA samples was hybridized with the microarray chips containing probes for the 1,200 mts-ipRNA.Hybridization images were acquired using a GenePix® 4000B laser scanner (Molecular Devices, Sunnyvale, CA) and digitized using Array-Pro® Analyzer image analysis software (Media Cybernetics, Rockville, MD). Relative signal values ​​were derived by background subtraction and normalization. Differentially expressed signals were determined using the ICON test (p < 0.05). From microarray analysis, approximately 500 of the 1,200 mts-ipRNAs were identified as highly specific for tassel. RQQnnn / ι ζπζ / β / υιλι TABLE 2 Description of the tissue samples used in the microarray assay Chip Number Genotype Tissue Type Pooled Samples from 3 Plants Stage 1 LH244 Youngest Ear <5 cm pooled VT 2 LH244 Oldest Ear >5 cm pooled VT-R1 3 LH244 Youngest Tassel 2-7 cm pooled V8-V9 4 LH244 Oldest Tassel >7 cm pooled V10-V12 5 LH244 Leaf V4 pooled V4 6 LH244 Leaf V12 pooled V12 7 LH244 Root V4 pooled V4 8 LH244 Stem V4 pooled V4 9 LH244 Youngest Ear <5 cm pooled VT 10 LH244 Oldest Ear >5 cm pooled VT-R1 11 LH244 Youngest Tassel 2-7 cm pooled V8-V9 12 LH244 Oldest Tassel >7 cm pooled V10-V12 13 LH244 leaf V4 clustered V4 14 LH244 leaf V12 clustered V12 15 LH244 root V4 clustered V4 16 LH244 root V4 clustered V4 17 01DKD2 Youngest ear <5 cm clustered VT 18 01DKD2 Oldest ear >5 cm clustered VT-R1 19 01DKD2 Oldest ear >5 cm clustered VT-R1 20 01DKD2 Youngest tassel 2-7 cm clustered V8-V9 21 01DKD2 Youngest tassel 2-7 cm clustered V8-V9 22 01DKD2 Oldest tassel >7 cm clustered V10-V12 23 01DKD2 Oldest tassel >7 cm clustered V10-V12 2401DKD2 Sheet V4 grouped V4 25 LH244 sheet V4 grouped V4 26 LH244 sheet V12 grouped V12 Bioinformatics analysis was then performed using sequence alignment tools such as the Basic Local Alignment Search Tool (BLAST) or SHort Read Mapping Package (SHRiMP) (Rumble, 2009) to compare the 500 mts-pRNA sequences identified as highly specific to tassel against a collection of unigene maize cDNA sequences. This BLAST analysis revealed maize cDNA sequences with which many mts-pRNA sequences aligned, yielding identifiable DNA sequence regions that have clustered, overlapping alignments of multiple mts-pRNA sequences with perfect or near-perfect matches. Six cDNA sequences were identified from this analysis as containing one or more of such mts-pRNA target sequence-rich regions. These six cDNA sequences are provided herein as SEQ ID NO: 17; SEQ ID NO: 18; SEQ ID NO: 19; SEQ ID NO: 20; SEQ ID NO: 21 and SEQ ID NO: 22.As an example, Figure 2 shows a graphical representation of the alignment of multiple mts-pRNA sequences in the cDNA provided as SEQ ID NO: 17. The multiple short lines represent the relative position of the mts-pRNA target sequences (and thus the location of the mts-pRNA binding site on the transcribed mRNA molecule) that align with the cDNA. The Y-axis represents the normalized mts-pRNA expression levels in male tissues as detected by microarray analysis. The box represents the cDNA region SEQ ID NO: 17 corresponding to the mts-pRNA target element sequences SEQ ID NO: 1 and SEQ ID NO: 2. Selected mts-pRNA sequences that align with one of the six cDNA sequences were used for further microarray analyses to determine differential expression in maize tissues. Microarray analyses of mts-pRNA sequences for normalized signal values ​​for the V8-V9 tassel, V10-V12 tassel, or combined signals from the other tissues for the LH244 and O1DKD2 maize germplasm are presented in Tables 3-10. Each table shows a subset of mts-pRNA sequences identified as aligning with one of the six cDNA sequences provided herein as SEQ ID NO: 17; SEQ ID NO: 18; SEQ ID NO: 19; SEQ ID NO: 20; SEQ ID NO: 21 and SEQ ID NO: 22. The results of the signal values ​​are measured as relative signal values ​​and the standard error (p<0.05) is represented by (STDR).The microarray results illustrate that representative mts-pRNA sequences show a high signal in the tassel (V8-V9 and V10-V12) and a low signal in other tissues, indicating that the endogenous expression of these mts-pRNAs is highly enriched in the tassel. The mts-pRNA sequence corresponds to the sense or antisense strand of the corresponding mts-pRNA target sequence in the cDNA sequence. The mts-pRNA sequence may have a single, two-nucleotide, or three-nucleotide mismatch with the aligned portion of the cDNA sequence, and the mts-pRNA sequence may align with the sense or antisense strand of the cDNA sequence. The mts-pRNA sequences are provided. RQonnn / i ζπζ / ε / υιλι are currently found in different maize germplasms. Table 3 shows relative microarray signal results for representative mts-pRNA sequences (provided herein as SEQ ID NO: 23–31) that align with the cDNA sequence provided herein as SEQ ID NO: 17, which contains the mts-pRNA target element sequences represented by SEQ ID NO: 1 and SEQ ID NO: 2. For the LH244 and O1DKD2 germplasms, the signals for these mts-pRNA sequences in the V8–V9 and V10–V12 tassels are higher than the signal for these mts-pRNA sequences in the other tissue samples. The microarray results shown in Table 3 indicate that the cDNA sequence provided herein as SEQ ID NO: 17 can be used as a source for designing mts-pRNA target elements for recombinant DNA molecules. RQonnn / i ζπζ / β / υιλι TABLE 3 SEO cDNA microarray results ID NO: 17 LH244 01DKD2 SEQ ID NO: V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) 23 183.5 (36.9) 514(105.2) 2.6 (0.6) 27 (2) 318 (46.5) 4.3(1.5) 24 104.5 (40.9) 322 (77.5) 7.7 (3.5) 16(1) 174 (32.3) 3 (0.7) 25 289 (99) 824.3 (146.6) 2.6 (0.7) 56 (9.1) 440 (14.1) 2.3(1.1) 26 55.5 (4.5) 175.7 (41.2) 3.8 (1.1) 10(7.1) 104 (26.3) 3.8(1.8) 27 377.5 (60.1) 581.7 (76.8) 1.1 (0.3) 73 (10.1) 405.5 (3.5) 1 (0.4) 28 126.5 (32.8) 248 (46.1) 2.4 (1.4) 29 (11.1) 134.5 (31.8) 3-8(1) 29 292.5 (25.8) 886.7 (99.7) 2.2 (0.7) 86 (11.1) 535.5 (20.7) 4(1-2) 30 173.5 (52) 495.7 (80.9) 4.2 (1.1) 32(1) 282 (20.2) 6.5(1.9) 31 8(3) 49 (15.8) 1.2 (0.4) 0(0) 26.5 (6.6) 2.8 (0.6) Table 4 shows relative microarray signal results for representative mts-pRNA sequences (provided herein as SEQ ID NO: 32–38) that align with the cDNA sequence provided herein as SEQ ID NO: 18, which contains the mts-pRNA target element sequence represented by SEQ ID NO: 3. For the LH244 germplasm, the signals for these mts-pRNA sequences in the V8–V9 and V10–V12 tassels are higher than the signals for these mts-pRNA sequences in the other tissue samples. For the O1DKD2 germplasm, the signals for these mts-pRNA sequences are higher in the V10–V12 tassel than the signals for these mts-pRNA sequences in the V8–V9 tassel or the other tissue samples. RQonnn / i ζηζ / Β / γίΛΐ TABLE 4 SEO cDNA microarray results ID NO: 18 LH244 01DKD2 SEQ ID NO: V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) 32 376.5 (320.7) 118 (30.8) 2 (0.6) 7.5 (3.5) 516 (307.1) 2.8 (0.9) 33 111.5 (12.6) 1032.7 (26.8) 17.8 (13.1) 4.5 (2.5) 648.5 (174.3) 2.8(1) 34 848.5 (675.3) 572.7 (211.3) 3 (0.6) 8.5 (0.5) 982 (389.9) 3.5(1.3) 35 437 (370.7) 117.7 (22.8) 1.6(0.6) 4(3) 519 (319.2) 2.8 (0.5) 36 906.5 (668.2) 606.7 (231.5) 2.2 (0.7) 15.5 (1.5) 1225.5 (480.3) 4.3(1.5) 37 554 (370.7) 553 (48.5) 1.5 (0.4) 9.5(1.5) 1224.5 (153) 3.3 (1.4) 38 768.5 (480.3) 1442.7 (17.2) 1.4(0.5) 8.5 (2.5) 1305.5 (10.6) 3(1.5) Table 5 shows relative microarray signal results for representative mts-pRNA sequences (provided herein as SEQ ID NO: 39-42) that align with the cDNA sequence provided herein as SEQ ID NO: 18, which contains the mts-pRNA target element sequence represented by SEQ ID NO: 4. For germplasm LH244, the signals for mts-pRNA sequences in the V8-V9 and V10-V12 tassels are higher than the signal for these mts-pRNA sequences in samples from other tissues. For germplasm 01DKD2, the signals for mts-pRNA sequences are higher in the V10-V12 tassel than the signal for these mts-pRNA sequences in the V8-V9 tassel or in samples from other tissues.The microarray results shown in Table 4 and Table 5 indicate that the cDNA sequence provided herein as SEQ ID NO: 18 can be used as a source for designing mts-ipRNA target elements for recombinant DNA molecules. Αοοηηη / ι ζάζ / β / υιλι TABLE 5 SEO cDNA microarray results ID NO: 18 LH244 01DKD2 SEQ ID NO: V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) 39 442.5 (242.9) 2157.3 (1059.3) 0.6 (0.2) 16 (12.1) 1514.5 (525.8) 1.5 (1.5) 40 119 (2) 448.7 (125.1) 1.5 (0.5) 7.5 (0.5) 452 (97) 2.8 (0.9) 41 185.5 (19.7) 702.7 (195.2) 0.9 (0.3) 5(1) 752 (225.3) 3(1.5) 42 143.5 (7.6) 564.3 (154.3) 2.6 (0.8) 6(1) 588 (163.6) 5.3 (1.5) Table 6 shows relative microarray signal results for representative mts-pRNA sequences (provided herein as SEQ ID NO: 43-46) that align with the cDNA sequence provided herein as SEQ ID NO: 19, which contains the mts-pRNA target element sequence represented by SEQ ID NO: 5. For LH244 germplasm, the signals for mts-pRNA sequences in the V8-V9 tuft and the V10-V12 tuft are higher than the signal for these mts-pRNA sequences for samples from other tissues. For germplasm 01DKD2, the signal for mts-pRNA sequences (SEQ ID NO: 43 - 44) is higher in the V10-V12 blob than the signal for these mts-pRNA sequences for the V8-V9 blob or other tissue samples; and the signals for mts-pRNA sequences (SEQ ID NO: 45 - 46) are higher than the signal for these mts-pRNA sequences in the V8-V9 blob and the V10-V12 blob compared to the other tissue sample. TABLE 6 SEO cDNA microarray results ID NO: 19 LH244 01DKD2 SEQ ID NO: V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) 43 52.5 (29.8) 147.3 (13.2) 1 (0.3) 4(0) 190.5 (2.5) 4.3 (2.4) 44 756 (548.5) 1340 (75) 1.8 (0.5) 15.5(1.5) 1443.5 (85.4) 6.3(1.7) 45 7901 (1803.1) 9982.3 (2999) 8.2 (5.4) 253 (11.1) 7195.5 (759.1) 5.8 (4.1) 46 4502.5 (424.8) 8663.3 (1595.4) 8.4 (6.2) 252 (9.1) 6778 (124.2) 6.8 (3.8) Table 7 shows relative microarray signal results for representative mts-pRNA sequences (provided herein as SEQ ID NO:47-52) that align with the cDNA sequence provided herein as SEQ ID NO:19, which contains the mts-pRNA target element sequence represented by SEQ ID NO:6. For the LH244 germplasm, the signals for the mts-pRNA sequences in the V8-V9 tuft and the V10-V12 tuft are higher than the signal for these mts-pRNA sequences for the other tissue samples.For germplasm 01DKD2, the signals for mts-pRNA sequences are higher in the V10-V12 blob than the signal for these mts-pRNA sequences in the V8-V9 blob or other tissue samples; the signals for mts-pRNA sequences (SEQ ID NO:47 and SEQ ID NO: 48) are moderately higher in the V8-V9 blob compared to the signal for these mts-pRNA sequences for the other tissue sample; the signal for mts-pRNA sequence (SEQ ID NO: 52) is significantly higher in the V8-V9 blob compared to the signal for this mts-pRNA sequence for the other tissue sample; and the signals for the mts-pRNA sequences (SEQ ID NO: 49, SEQ ID NO: 50 and SEQ ID NO: 51) are not significantly different from the signal for these mts-pRNA sequences for samples from other tissues.The microarray results shown in Table 6 and Table 7 indicate that the cDNA sequence provided herein as SEQ ID NO: 19 can be used as a source for designing mts-ipRNA target elements for recombinant DNA molecules. Αοοηηη / ι ζπζ / ε / υιλι TABLE 7 SEO cDNA microarray results ID NO: 19 LH244 01DKD2 SEQ ID NO: V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) 47 157.5 (5.6) 900.7 (51.8) 15.5 (7.4) 12.5 (1.5) 1051.5 (199.5) 4 (0.7) 48 221 (106.1) 720.3 (251.7) 4.8 (2.7) 9.5 (6.6) 470.5 (57.1) 1 (0.4) 49 340 (239.4) 1241.7 (105.7) 16.3 (5.4) 57.5 (10.6) 867 (117.2) 55.3 (11.6) 50 296 (228.3) 722 (267.2) 0(0) 5.5(5.6) 326 (262.6) 1.8(1.8) 51 158 (78.8) 287 (46) 34.3 (6.5) 24.5 (0.5) 275.5 (20.7) 40 (3.1) 52 4080 (1829.4) 3629.7 (1327.5) 5.5(3.5) 234 (11.1) 3286.5 (33.8) 3.5 (2.1) Table 8 shows relative microarray signal results for representative mts-pRNA sequences (provided herein as SEQ ID NO: 53–60) that align with the cDNA sequence provided herein as SEQ ID NO: 20, which contains the mts-pRNA target element sequence represented by SEQ ID NO: 7. For the LH244 and O1DKD2 germplasms, the signals for these mts-pRNA sequences in the V8V9 and V10–V12 tassels are higher than the signal for these mts-pRNA sequences in the other tissue samples. The microarray results shown in Table 8 indicate that the cDNA sequence provided herein as SEQ ID NO: 20 can be used as a source for designing mts-pRNA target elements for recombinant DNA molecules. RQQnnn / ι ζπζ / β / υιλι TABLE 8 SEO ID NO: 20 cDNA microarray results LH244 01DKD2 SEQ ID NO: V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) 53 76.5 (1.5) 145 (19.4) 2.2 (0.8) 16 (2) 84(1) 1.5(0.5) 54 100.5 (3.5) 172 (21.6) 14.2 (6.2) 56 (22.2) 231 (79.8) 19.5 (4.7) 55 377.5 (60.1) 581.7 (76.8) 1.1 (0.3) 73 (10.1) 405.5 (3.5) 1 (0.4) 56 261 (1) 692 (59.7) 49.5 (30.4) 138.5 (29.8) 599.5 (41.9) 19.3 (4.8) 57 126.5 (32.8) 248 (46.1) 2.4(1.4) 29 (11.1) 134.5 (31.8) 3.8(1) 58 215.5 (14.6) 349 (29.1) 0.7 (0.3) 88.5 (2.5) 327 (31.3) 2 (0.9) 59 789 (97) 1262.7 (169.2) 4.1 (2.6) 202 (24.2) 811.5 (115.7) 0.3 (0.3) 60 141.5 (9.6) 265.7 (30.1) 5.8(1.2) 75.5 (5.6) 303 (37.4) 9.3(1.8) Table 9 shows relative microarray signal results for representative mts-pRNA sequences (provided herein as SEQ ID NO: 61–69) that align with the cDNA sequence provided herein as SEQ ID NO: 21, which contains the mts-pRNA target element sequence represented by SEQ ID NO: 8. For the LH244 and O1DKD2 germplasms, the signals for the mts-pRNA sequences in the V8–V9 and V10–V12 tassels are higher than the signal for these mts-pRNA sequences in the other tissue samples. The microarray results shown in Table 9 indicate that the cDNA sequence provided herein as SEQ ID NO: 21 can be used as a source for designing mts-pRNA target elements for recombinant DNA molecules. RQonnn / i zciz / r / yiai TABLE 9 SEO cDNA microarray results ID NO: 21 LH244 01DKD2 SEQ ID NO: V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) V8-V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) 61 850.5 (8.6) 334.7 (222.3) 9.4 (3.4) 461 (154.6) 454 (1) 8(5) 62 44.5 (5.6) 11.3(3.9) 1.8(0.7) 13.5 (5.6) 15.5 (6.6) 1(1) 63 34.5 (4.5) 11.7(2.2) 3.2(1) 14.5 (1.5) 18 (1) 1.5 (0.5) 64 23 (8.1) 13.7 (4.6) 1.8(1) 13.5 (13.6) 9 (8.1) 1.3 (0.9) 65 811.5 (169.2) 260.3 (103.2) 5.8 (2.7) 444.5 (4.5) 454.5 (45) 12 (8.4) 66 199.5 (54) 48 (20.6) 1.9 (0.7) 61 (44.4) 55.5 (34.9) 2.8(1.8) 67 216 (18.2) 62 (16.6) 3.3 (2) 123.5 (0.5) 107 (17.2) 4.3 (2.6) 68 265 (86.9) 85.3 (20.4) 5 (2.3) 98 (62.6) 96 (56.6) 2.5 (2.2) 69 516 (82.8) 185 (82.1) 11.2(4.1) 309.5 (19.7) 282.5 (2.5) 9.8(5.5) Table 10 shows relative microarray signal results for representative mts-pRNA sequences (provided herein as SEQ ID NO: 70-87) that align with the cDNA sequence provided herein as SEQ ID NO: 22, which contains the mts-pRNA target element sequence represented by SEQ ID NO: 9. For LH244 germplasm, the signals for the mts-pRNA sequences (SEQ ID NO: 70, 71, 73, 75-81, 83-87) in the V8-V9 tassel and the V10-V12 tassel are higher than the signal for these mts-pRNA sequences for samples from other tissues; and the signals for mts-ipRNA sequences (SEQ ID NO: 72, 74, 82) in other tissue were higher than the signal for these mts-ipRNA sequences in the V8V9 blob or the V10-V12 blob.For germplasm 01DKD2, the signals for mtsnRNA sequences (SEQ ID NO: 70-87) are higher in the V10-V12 tuft than the signal for these mtsnRNA sequences in the V8-V9 tuft or other tissue samples. The microarray results shown in Table 10 indicate that the cDNA sequence provided herein as SEQ ID NO: 22 can be used as a source for designing mtsnRNA target elements for recombinant DNA molecules. RQonnn / i zciz / r / yiai TABLE 10 SEO cDNA microarray results ID NO: 22 LH244 01DKD2 SEQ ID NO: V8V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) V8V9 Tassel (STDR) V10-V12 Tassel (STDR) Other Tissue (STDR) 70 56.5 (23.7) 694 (256.9) 1.8 (0.4) 3(3) 873 (308.1) 4(1.5) 71 1125.5 (383.4) 2380 (696.9) 1.6 (0.5) 46 (0) 1943.5 (447) 2.3(1.3) 72 323.5 (54) 325.7 (55.9) 1324.8 (1083.3) 332.5 (83.3) 530 (89.9) 233.3 (16.5) 73 1058.5 (370.2) 2395.7 (634.9) 4.5 (1.8) 37.5 (2.5) 1695.5 (380.3) 2 (1.4) 74 83.5 (20.7) 129 (11.2) 164.2 (106.5) 43.5 (0.5) 171 (19.2) 45.8 (4.5) 75 53 (34.3) 237.3 (62.1) 3.3 (0.5) 3(1) 257 (74.8) 5(1.8) 76 55 (38.4) 252.3 (66.5) 5.7(1) 2.5 (0.5) 276.5 (81.3) 4(1.4) 77 180.5 (64.1) 302.7 (111.2) 4.1 (2.3) 0.5 (0.5) 225 (30.3) 2.5 (0.6) 78 47.5 (15.7) 642.7 (242) 1 (0.4) 3(2) 715.5 (211.6) 4.5(1) 79 197.5 (62.1) 269.3 (106.8) 1.9 (0.6) 5(1) 290.5 (28.8) 2 (0.4) 80 138 (83.8) 383.3 (44.6) 0.7 (0.3) 2.5(1.5) 230 (43.4) 0.5 (0.3) 81 207 (113.1) 470.3 (55.2) 1.6(1.1) 1(1) 320 (69.7) 0(0) 82 95.5 (29.8) 137 (17.7) 217.4 (158.6) 71.5(7.6) 211 (14.1) 54.8 (5.1) 83 173 (63.6) 303(82) 2.5 (1.2) 2(1) 253.5 (24.7) 1.5(0.6) 84 13 (5.1) 35.7 (7.1) 4.4 (1.5) 0(0) 20(10.1) 1 (0.4) 85 184 (71.7) 310(109.1) 2.8(1.3) 2.5 (0.5) 246 (14.1) 2.5(1) 86 173.5 (62.1) 241.7 (101.2) 2.4 (0.7) 3(1) 256.5 (7.6) 3 (0.9) 87 56 (35.4) 104 (12.7) 4.7 (2) 2.5 (0.5) 91.5 (6.6) 2.5(1). These analyses confirmed that the six cDNA sequences (SEQ ID NO: 17–22) were rich in mts-pRNA target sequences and that the corresponding mts-pRNA showed high tassel specificity. Therefore, these cDNA sequences contain sequences useful for molecular biology methods to create recombinant DNA molecules containing mts-pRNA target elements that can confer mts-pRNA-mediated transgene silencing. An analysis of the corresponding genomic sequence containing the mts-siRNA target elements was performed for thirty-two different maize germplasms (with a relative maturity (RM) in the range of 80 to 120 days), typically used as a female in a hybrid cross, to confirm the presence and any sequence variation of the mts-siRNA target elements. For the mts-siRNA target elements provided as SEQ ID NO: 1 and SEQ ID NO: 2, three sets of thermal amplification primer pairs were designed to amplify the corresponding sequence within the cDNA sequence provided herein as SEQ ID NO: 17. These primers were used to generate a PCR amplicon in the genomic DNA extracted from the tissue of each germplasm. Amplicons were produced from all the evaluated germplasms.The amplicon sequence in the thirty-two germplasms was either 100% identical to the mts-pRNA target element sequence or contained a minimal amount of single nucleotide polymorphisms (up to 95% identity). These data indicate that transgenic plants generated with a recombinant DNA construct comprising a transgene encoding a recombinant protein operatively linked to an mts-pRNA target element provided as SEQ ID NO: 1 or SEQ ID NO: 2 would exhibit male tissue-specific regulation of recombinant protein expression in most maize germplasms. If the transgene encodes a recombinant protein conferring glyphosate tolerance, then tassels in most maize germplasms would exhibit glyphosate-induced male sterility. EXAMPLE 2 recombinant DNA constructs vs plant transformation vectors Recombinant DNA constructs and plant transformation vectors were created using DNA sequences corresponding to regions of the six cDNA sequences identified as rich in mts-pRNA target sequences. The recombinant DNA constructs and plant transformation vectors were designed to be useful for producing transgenic plants in which tassel-specific silencing of a transgene operatively linked to an mts-pRNA target element is achieved through mts-pRNA-mediated silencing. Nine mts-pRNA target elements were designed using the results of the analyses of the six cDNA sequences. Each mts-pRNA target element was designed to have a DNA sequence comprising many overlapping mts-pRNA target sequences to which different mts-pRNAs can bind. The mts-pRNA target element sequence provided herein as SEQ ID NO: 1 has 95% sequence identity with nucleotide positions 1429 to 1628 of the cDNA sequence provided herein as SEQ ID NO: 17. The mts-pRNA target element sequence provided herein as RQQnnn / ι ζπζ / β / υιλι SEQ ID NO: 2 has a single nt change (T69A) with respect to SEQ ID NO: 1. The mts-pRNA target element sequence provided herein as SEQ ID NO: 3 corresponds to nucleotide positions 239 to 433 of the cDNA sequence provided herein as SEQ ID NO: 18. The mts-pRNA target element sequence provided herein as SEQ ID NO: 4 corresponds to nucleotide positions 477 to 697 of the cDNA sequence provided herein as SEQ ID NO: 18. The mts-pRNA target element sequence provided herein as SEQ ID NO: 5 corresponds to nucleotide positions 239 to 433 of the cDNA sequence provided herein as SEQ ID NO: 19. The mts-pRNA target element sequence provided herein as SEQ ID NO: 6 corresponds to the nucleotide position 370 to 477 of the cDNA sequence provided herein as SEQ ID NO: 19.The mts-pRNA target element sequence provided herein as SEQ ID NO: 7 corresponds to nucleotide positions 1357 to 1562 of the cDNA sequence provided herein as SEQ ID NO: 20. The mts-pRNA target element sequence provided herein as SEQ ID NO: 8 corresponds to nucleotide positions 247 to 441 of the cDNA sequence provided herein as SEQ ID NO: 21. The reverse complement of the mts-pRNA target element sequence represented by SEQ ID NO: 9 has 99% sequence identity with nucleotide positions 191 to 490 of the cDNA sequence provided herein as SEQ ID NO: 22, with three nt mismatches (C314A, A350G, and G408A) of SEQ ID NO: 22 with respect to the SEQ reverse complement sequence ID NO: 9.Other mts-pRNA target elements can be created by combining the DNA sequences of different mts-pRNA target elements or fragments of two or more cDNA regions rich in mts-pRNA target elements, for example, by combining all or fragments of two or more of the mts-pRNA target elements provided herein as SEQ ID NO: 1-9 and / or one or more of the mts-pRNA sequences provided herein as SEQ ID NO: 23-92. Different fragments of these mts-pRNA target elements ranging in length from 21 to 170 nucleotides were combined and new mts-pRNA target elements were produced using DNA synthesis methods known to the art and are provided as SEQ ID NO: 10-16. The mts-pRNA target elements were subcloned into recombinant DNA constructs with the mts-pRNA target element operatively linked to the 3' end of the open reading frame of the cp4-epsps gene (SEQ ID NO: 93), which encodes the CP4-EPSPS protein for glyphosate tolerance, and 5' to the 3' operatively linked untranslated region (3'-UTR). The recombinant DNA constructs also contained operatively linked combinations of one of three different promoter / intron / enhancer combinations and a chloroplast transit peptide sequence. The expression cassette configurations contained in the vectors RQQnnn / ι ζπζ / β / υιλι transformation to evaluate the targeting efficiency of mts-siRNA. EXAMPLE 3 plant transformation vs efficacy tests Transformation vectors containing recombinant DNA constructs were used with an Agrobacterium-based transformation method and immature maize embryos, following established procedures. Leaf samples were collected from R0 plants and subjected to molecular analysis to select transgenic plants containing either a single copy (one insertion) or a double copy (two independent insertions) of the recombinant DNA construct and lacking vector structure. Single-copy plants were not sprayed with glyphosate, were self-pollinated, and advanced to R1 seed collection, while double-copy plants were sprayed with glyphosate to evaluate vegetative tolerance and induced male sterility.Transgenic plants that were not sprayed with glyphosate had normal anthesis, normal pollen dispersal, and normal pollen development as determined by Alexandar staining and microscopic observation. Double-copy R0 events were used in the evaluations to calculate a single-copy homozygous event. Plants were sprayed with glyphosate at two different growth stages to determine whether the presence of the mts-ipRNA target element in the recombinant DNA construct confers vegetative glyphosate tolerance and glyphosate-induced tassel sterility. Plants were sprayed in a greenhouse with 100% glyphosate (0.75 Lb ae / acre) applied at the V5 stage, followed by 100% glyphosate (0.75 Lb ae / acre) applied at the V8 stage. Seven days after the V5 glyphosate application, plants were scored for vegetative lesions, with lesion scores of <10% considered indicative of vegetative glyphosate tolerance (% vegetative tolerance). Male sterility was measured in two ways.Plants exhibiting vegetative glyphosate tolerance were scored after V8-stage glyphosate application for complete glyphosate-induced male sterility, measured as the absence of anther extrusion up to stage S90 + 12 (% complete male sterility). Anthers were collected and dissected for microscopic observation of pollen development. For each event, anthers were assessed for non-viable pollen grains produced, as detected by Alexandar staining under microscopic observation, and scored as producing no viable pollen (% no pollen). In plants not sprayed with glyphosate, tassel development, anther extrusion, and pollen development were normal. RQonnn / i zciz / r / yiai The target elements of mts-ipRNA were evaluated in multiple transformation vector configurations. Each transformation vector was used to produce multiple R0 plants, with each R0 plant representing a unique transgenic event created using the same transformation vector. Data for three different transformation vector configurations are provided in Tables 11 and 12. Twelve mts-ipRNA target elements were evaluated in a first vector configuration (A) with the results shown in Table 11. Surprisingly, the percentage of plants showing vegetative glyphosate tolerance ranged from 80-100%, but the percentage showing complete glyphosate-induced male sterility (induced male sterility included non-viable pollen and absence of pollen production) ranged from 0-100%. See Table 11. For example, plants produced using a transformation vector containing the mts-ipRNA target element encoded by SEQ ID NO: 13, SEQ ID NO: 14, or SEQ ID NO: 15 had high amounts showing vegetative glyphosate tolerance (in the range of 90-100% of the plants evaluated), but none of the transgenic events showed complete glyphosate-induced male sterility;Plants produced using a transformation vector containing the mts-ipRNA target element encoded by SEQ ID NO: 5 or SEQ ID NO: 9 had high amounts showing vegetative glyphosate tolerance (in the range of 90-100% of the plants evaluated) and moderate amounts showing glyphosate-induced male sterility (in the range of 50-40% of the plants evaluated);Plants produced using a transformation vector containing the mts-ipRNA target element encoded by SEQ ID NO: 2, SEQ ID NO: 7, or SEQ ID NO: 8 had high amounts of plants exhibiting vegetative glyphosate tolerance (88–100% of evaluated plants) and high amounts of plants exhibiting glyphosate-induced male sterility (82–100% of evaluated plants). Pollen development was assessed using Alexandar staining. Plants containing four events created with the transformation vector containing the mts-ipRNA target element provided as SEQ ID NO: 2 and sprayed with glyphosate to induce male sterility showed very few aborted pollen grains or no pollen grains were detected. Plants that did not receive glyphosate application had normal pollen. See Figure 4. RQonnn / i zciz / r / yiai AQQnnn / i ζπζ / ε / υιλι TABLE 11 RO plant evaluation SEQ ID NO: Vector configuration Number of plants sprayed % of vegetative tolerance % of complete male sterility % of no pollen 2 A 11 100 82 82 3 A 10 90 10 0 4 A 7 86 7 0 5 A 10 90 50 0 6 A 9 90 82 0 7 A 9 88 100 44 8 A 10 90 100 50 9 A 10 100 40 0 ​​13 A 7 100 0 0 14 A 10 90 0 0 15 A 10 90 0 0 16 A 10 80 50 0 Four additional mts-ipRNA target elements were evaluated in a second and third vector configuration (B and C, respectively), with the results shown in Table 12. Differences in the percentage of plants exhibiting vegetative glyphosate tolerance and the percentage exhibiting complete glyphosate-induced male sterility were observed between the two vector configurations. Vector configuration B yielded an increased percentage of plants exhibiting vegetative glyphosate tolerance for all evaluated mts-ipRNA target elements.For the percentage that showed complete glyphosate-induced male sterility, two of the mts-siRNA target elements (SEQ ID NO: 10 and SEQ ID NO: 11) evaluated had better results in the C vector configuration, one of the mts-siRNA target elements (SEQ ID NO: 12) had better results in the B vector configuration, and one of the mts-siRNA target elements (SEQ ID NO: 1) provided 100% complete glyphosate-induced male sterility in the B and C configurations. TABLE 12 RO plant evaluation SEQ ID NO: Vector configuration Sprayed plants % of vegetative tolerance % of complete male sterility % of no pollen 1 B 9 67 100 100 10 B 10 90 55 22 11 B 2 100 50 0 12 B 10 100 50 20 1 C 13 38 100 60 10 C 6 83 100 0 11 C 9 55 80 0 12 C 15 87 23 0 RQonnn / i ζπζ / β / υ Of the plants exhibiting vegetative glyphosate tolerance, the percentage of glyphosate-induced male-sterile plants lacking viable pollen ranged from 0 to 100%. For plants produced using a transformation vector that had more than 60% of evaluated plants exhibiting both vegetative glyphosate tolerance and complete glyphosate-induced male sterility, the percentage of plants lacking viable pollen ranged from 0 to 100%. Two mts-siRNA target elements (SEQ ID NO: 1 in vector configuration B and SEQ ID NO: 2 in vector configuration A) had 100% and 82% lacking viable pollen, respectively. These two mts-siRNA target elements (SEQ ID NO: 1 and SEQ ID NO: 2) differ by one nucleotide and are derived from the same cDNA sequence (SEQ ID NO: 17). EXAMPLE 4 Immunolocalization of CP4-EPSPS orotein in tassel Immunolocalization of CP4-EPSPS protein in pollen tassels from transgenic plants was used to analyze protein expression at the cell and tissue levels to confirm the loss of CP4-EPSPS protein expression in pollen due to the presence of an operatively bound mts-ipRNA target element. R3 generation transgenic plants containing the cp4-epsps transgene operatively bound to SEQ ID NO: 1, or as a control, the cp4-epsps transgene without an operatively bound mts-ipRNA target element, were grown in a greenhouse. Plants were sprayed with glyphosate IX (0.75 lb ae / acre) at the V2 stage to confirm vegetative tolerance. Pollen tassels were harvested at 1 cm to 17 cm in length at the V8 to V12 stages when the anther tissue was in the microspore stem cell and free microspore stages.The anthers were removed from the tassel spike using dissecting forceps and immediately fixed in 3.7% formaldehyde in phosphate-buffered saline (PBS) under gentle vacuum. After washing in PBS, the tissues were placed in embedding medium and immediately frozen. The frozen tissue blocks were stored at -80 °C until sectioned in a microtome at -20 °C and collected on loaded slides. Tissue sections were blocked with blocking agent (10% normal goat serum, 5% bovine serum albumin, 0.1% Triton X-100 in PBS) for two hours. The sections were incubated with anti-CP4-EPSPS antibody (1 / 500 in PBS). After washing the sections three times in PBS, tissue sections were incubated with the secondary antibody, Alexa Flour® 488 conjugated anti-goat mouse IgG (Invitrogen, Eugene, Oregon). A negative control was prepared by omitting the CP4-EPSPS antibody incubation. The primary and secondary antibodies were incubated at room temperature for two to four hours and then further incubated overnight at 4°C. After washing, images of the tissues were obtained using a Zeiss META 510 confocal Laser Scanning Microscope (LSM) using a 488 nm laser for excitation and 500-550 nm (green channel) for the emission filter setting.The same imaging parameters were applied to the samples, including controls. Bright-field and fluorescent images of each section were scanned and then fused using LSM software to display structural information. Data for the negative controls showed the expected absence of signal. Data for the transgenic plants containing the cp4-epsps transgene operatively linked to SEQ ID NO: 1 showed a low fluorescence signal, indicating low CP4-EPSPS protein expression in the anther wall, tapetum, and developing pollen microspores of the anther. Data for the control transgenic plants (those containing the cp4-epsps transgene operatively linked to an mts-ipRNA target element) showed a high fluorescence signal, indicating high CP4-EPSPS protein expression in the anther wall, tapetum, and developing pollen microspores of the anther. The loss of CP4-EPSPS protein expression in pollen from plants containing the cp4-epsps transgene operatively linked to the mts-ipRNA target element provided as SEQ ID NO: 1 correlates with the complete glyphosate-induced male sterility observed in these plants. These data confirmed that the observed complete glyphosate-induced male sterility results from the loss of CP4-EPSPS protein expression in pollen due to the presence of the operatively linked mts-ipRNA target element provided as SEQ ID NO: 1. EXAMPLE 5 Field trials For optimal use in hybrid production, very low anther extrusion under field conditions after herbicide application, combined with tolerance to vegetative herbicides (as measured by low crop damage), is desirable. Other aspects of hybrid maize production, such as plant height and yield, may also be desirable. To evaluate these, transgenic plants comprising the cp4-epsps transgene operatively linked to an mts-ipRNA target element were assessed in advanced generations under field conditions. RQonnn / i ζοζ / β / υιλι field and multiple parameters were measured. Field trials were conducted on R3 generation plants containing the cp4-epsps transgene operatively linked to the mts-ipRNA target element provided as SEQ ID NO: 1 at multiple locations to evaluate vegetative glyphosate tolerance and glyphosate sensitivity specific to tassel. The field trials evaluated R3 generation plants containing the same transgenic insert at different genomic locations. The plants evaluated contained a single copy of one of four unique transgenic events created using the same plant transformation vector containing the cp4-epsps transgene operatively linked to the mts-ipRNA target element provided as SEQ ID NO: 1. The trials were conducted using a randomized complete block design. Multiple agronomic and trait efficacy parameters were scored during the field trial season, and yield was determined at the end of the season.Trait efficacy field trials were conducted by applying glyphosate herbicide at 0.75 Lb ae / acre at the V7 stage, followed by 0.75 Lb ae / acre at V9, and assessing the percentage of crop injury at the VT stage (CIPVT), the average percentage of anther extrusion at the S90+8 stage (AES9E), and yield (measured as bushels per acre (bu / acre)) at the end of the season. The field trials included the glyphosate-tolerant transgenic event NK603 (ATCC deposit number PTA-24780) as a negative control for glyphosate-induced male sterility and as a positive control for vegetative glyphosate tolerance. Plants containing the NK603 event exhibit commercial-level vegetative glyphosate tolerance and produce fully glyphosate-tolerant tassels. All data were subjected to separate analysis of variance and mean (LSD) at p < 0.05. TABLE 13 Efficacy results of rasaos for field trials with plants containing SEO ID NO: 1 AQonnn / ι ζπζ / β / υιλι CIPVT event AES9E NK603 control 1.25 95.00 Event 1 1.25 0 Event 2 3.75 3.25 Event 3 0 4.00 Event 4 0 1.75 The average crop injury at the VT stage for control plants containing the NK603 event was 1.25. The average crop injury for plants containing event 1, event 2, event 3, and event 4 was 1.25, 3.75, 0, and 0, respectively. The least significant difference (LSD) in crop injury at the 0.05 level was 10.25 for all events evaluated. These results indicate that plants containing the four transgenic events with the mts-siRNA SEQ ID NO: 1 target element had no or very low vegetative injury with the application of 0.75 Lb ae / acre of glyphosate at V7, followed by V9, similar to the control plants containing the NK603 event. The average anther extrusion at stage S90+8 for the control plants containing the NK603 event was 95. The average anther extrusion at S90+8 for the plants containing event 1, event 2, event 3, and event 4 was 0, 3.25, 4, and 1.75, respectively. The average anther extrusion at S90+8 LSD at 0.05 was 42.71 for all events evaluated. These results indicate that plants containing the four transgenic events containing the mts-RNApi SEQ ID NO: 1 target element had zero or very low anther extrusion with the application of 0.75 Ib ae / acre of glyphosate at V7, followed by V9, contrary to the control plants containing the NK603 event, which were found to be completely male-fertile after the application of glyphosate. At the end of the season, the corn from these field trials was harvested and the yield was determined. The average yield for the control plants containing the NK603 event was 96.74 bu / acre. The average yield for the plants containing event 1, event 2, event 3, and event 4 was 102.17 bu / acre, 96.48 bu / acre, 97.59 bu / acre, and 95.8 bu / acre, respectively. The LSD of the yield at 0.05 was 26.25 bu / acre for all events evaluated. See Table 14. These results indicate that the plants containing the four transgenic events that contained the mts-IPRNA SEQ ID NO: 1 target element had yield parity with NK603. TABLE 14 Performance results for field trials with plants containing SEO ID NO: 1 AQonnn / ι ζπζ / ε / υιλι Event Average Yield (bu / acre) NK603 Control 96.74 Event 1 102.17 Event 2 96.48 Event 3 97.59 Event 4 95.8 Field trials were conducted on R2 or later inbred plants containing the cp4-epsps transgene operatively linked to various mts-ipRNA target elements at multiple locations to evaluate vegetative glyphosate tolerance and glyphosate sensitivity specific to tassel. The field trials evaluated R2 or later inbred plants containing a single copy of the cp4-epsps transgene operatively linked to the mts-ipRNA target element SEQ ID NO: 1, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8, each in the transformation vector configuration A. The trials were conducted using a cluster block design with the clustering factor being the transformation vector or a pool of transformation vectors if the event numbers for a specific transformation vector were low.Multiple agronomic and trait efficacy parameters were scored during the field trial season, and yield was determined at the end of the season. Trait efficacy field trials were conducted by applying glyphosate at 1.5 Lb ae / acre to V2 corn, followed by glyphosate at 0.75 Lb ae / acre to V8 corn (875 degree-of-growth days), followed by glyphosate at 0.75 Lb ae / acre to VIO corn (1025 degree-of-growth days). Scores were recorded for percentage of crop injury at the VT stage (CIPVT), average percentage of anther extrusion at the S90+8 stage (AES90+8), average plant height (in inches), and yield (measured as bushels per acre [bu / acre]) at the end of the season. Field trials included the glyphosate-tolerant transgenic event NK603 as a negative control for glyphosate-induced male sterility and as a positive control for vegetative tolerance to glyphosate.A glyphosate-tolerant mixture of male pollinators, consisting of three hybrid germplasm bases, was placed every three plots and surrounding the entire trial to serve as the pollen source for the test entries. Herbicide treatments were applied using a CO2 backpack sprayer or a suspended sprayer calibrated to deliver 15 gallons per acre (GPA) using TeeJet® TTI air-induced nozzles (TeeJet Technologies, Springfield, IL) with water as the herbicide carrier. All data were subjected to separate analysis of variance and mean square (LSD) at p < 0.05. The results are provided in Table 15. No significant difference was observed in the percentage of crop lesion in VT (CIPVT) or in average plant height compared to the NK603 control for the evaluated plants containing the mts-RNAi target element SEQ ID NO: 1, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, or SEQ ID NO: 8. Plants containing the mts-RNAi target element SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 6, or SEQ ID NO: 7 also did not have a significant decrease in seed yield compared to the NK603 control. Plants containing the mts-RNAi target element SEQ ID NO: 1 or SEQ ID NO: 7 had very low or absent anther extrusion with AES90+8 values ​​of 0–2.75% and 0–1.5%, respectively. The plants that contained the target element of mts-ARNip SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6 RQonnn / i 7Π7 / β / υιλι o SEQ ID NO: 8 had significantly decreased anther extrusion compared to control plants, with AES90+8 values ​​in the range of 10% to 25%. RQonnn / i ζηζ / Β / γίΛΐ TABLE 15 mts-ARNip different in the same configuration A of the transformation vector SEQ ID NO: Number of events evaluated CIPVT (LSD=8.1) AES90+8 (LSD = 10) Seed yield bu / acre (LSD=24.7) Average plant height (inches) (LSD=6.9) n / a NK603 0-6.25 % 90% 110 83.2 1 8 2.5-6.25 % 0-2.75 % 97-99 84.75 4 4 1.25- 3.75 % 16.25- 18.75 % 70-80 85.1 5 4 5 % 10 %- 20% 90-100 84.31 6 7 3.75 % 25% 120 82.1 7 2 3.75-5 % 0-1.5% 100-110 79.8 8 2 2.5-2.7 % 15.75- 17.85 % 80-90 80.1 EXAMPLE 6 Hybrid seed production The transgenic plants and seeds of the invention can be used for cultivation purposes, including in the production of hybrid seeds. The transgenic maize plants, comprising a recombinant DNA construct with a transgene encoding a glyphosate-tolerant EPSPS protein operatively linked to an mts-ipRNA target element, are planted in an area, for example, an open field. Other original maize plants may or may not be present in the same area. For weed control during seed production, glyphosate can be applied to the transgenic maize plants at the vegetative stages as directed on Roundup® agricultural product labels. Hybrid seed production can be achieved by applying glyphosate to transgenic (female) maize plants, starting just before or during tassel development at the V7 to V13 vegetative growth stages. Glyphosate application will induce a male-sterile phenotype through tissue-selective glyphosate tolerance in the transgenic maize plants. These induced male-sterile transgenic maize plants can then be pollinated by other pollen-donating (male) plants, resulting in viable hybrid maize seed containing the recombinant DNA construct for tissue-selective glyphosate tolerance. The pollen-donating plants may or may not be present in the same area and may or may not be transgenic maize plants. Pollination can be achieved through methods known in the art, including close proximity of plants or by hand pollination.The hybrid seed is harvested from the 5 transgenic corn plants. Having illustrated and described the principles of the present invention, it should be evident to those skilled in the art that the arrangement and details of the invention may be modified without departing from such principles. We claim that all modifications are within the spirit and scope of the appended claims.

Claims

NOVELTY OF THE INVENTION CLAIMS 1. A recombinant DNA molecule comprising an mts-siRNA target element comprising a sequence selected from the group consisting of SEQ ID NO: 7, 8, and complements thereof wherein the mts-siRNA target element is operatively linked to a heterologous polynucleotide molecule.

2. The recombinant DNA molecule according to claim 1, further characterized in that the heterologous transcribable polynucleotide molecule encodes a protein that confers herbicide tolerance in plants.

3. The recombinant DNA molecule according to claim 2, further characterized in that said heterologous transcribable polynucleotide molecule encodes a glyphosate-tolerant 5-enolipyruvyl shikimate 3-phosphate synthase (EPSPS).

4. A method for producing a recombinant DNA molecule, characterized in that it comprises operationally attaching an mts-pRNA target element to a heterologous transcribable polynucleotide molecule.

5. The method according to claim 4, further characterized in that said mts-RNAi target element comprises a sequence selected from the group consisting of SEQ ID NO: 7, 8, and complements thereof.

6. A transgenic plant or part thereof, characterized in that its genome comprises the recombinant DNA molecule as claimed in claim 1.

7. A seed of the transgenic plant as claimed in claim 6, characterized in that it comprises said DNA molecule.

8. The plant according to claim 6, further characterized in that said plant is a monocotyledonous plant.

9. The plant according to claim 8, further characterized in that said plant is a corn plant.

10. A method for selectively regulating the expression of a protein in a male reproductive tissue of a transgenic plant, characterized in that it comprises expressing in said transgenic plant the recombinant DNA molecule as claimed in claim 1.

11. The method according to claim 10, further characterized in that said protein comprises a glyphosate-tolerant 5-enolipyruvyl shikimate 3-phosphate synthase (EPSPS).

12. A method for inducing male sterility in a transgenic plant, characterized in that it comprises: a) cultivating a transgenic plant comprising a recombinant DNA molecule comprising an mts-pRNA target element comprising a sequence selected from the group consisting of SEQ ID NO: 7, 8, and complements thereof, wherein the mts-pRNA target element is operatively linked to a heterologous polynucleotide molecule encoding a protein conferring tolerance to at least one first herbicide; and b) applying an effective amount of said herbicide to said transgenic plant, wherein the herbicide application is carried out before or simultaneously with the development of the male reproductive tissue of said transgenic plant, thereby inducing male sterility in the transgenic plant.

13. The method according to claim 12, further characterized in that said heterologous transcribable polynucleotide molecule encodes a glyphosate-tolerant 5-enolipyruvil shikimate 3-phosphate synthase (EPSPS).

14. The method according to claim 12, further characterized in that said herbicide is glyphosate.

15. The method according to claim 14, further characterized in that said effective amount of herbicide is between approximately 0.125 pounds of acid equivalent per acre and approximately 8 pounds of acid equivalent per acre of glyphosate.

16. The method according to claim 12, further characterized in that said effective quantity of herbicide is applied at a developmental stage selected from the group consisting of stages V4, V5, V6, V7, V8, V9, V10, V11, V12, V13 and V14.

17. A method for producing a hybrid seed, characterized in that it comprises: a) applying an effective amount of herbicide to a transgenic plant comprising a recombinant DNA molecule comprising an mts-pRNA target element comprising a sequence selected from the group consisting of SEQ ID NO: 7, 8, and complements thereof, wherein the mts-pRNA target element is operatively linked to a heterologous transcribable polynucleotide molecule encoding a protein conferring tolerance to at least a first herbicide, wherein said herbicide application is carried out before or simultaneously with the development of the male reproductive tissue of the transgenic plant, thereby inducing male sterility in said transgenic plant; b) fertilizing said transgenic plant with pollen from a second plant; and c) allowing a hybrid seed to form from said transgenic plant.

18. The method according to claim 17, further characterized in that said fertilization comprises allowing wind pollination to occur.

19. The method according to claim 17, further characterized in that said heterologous transcribable polynucleotide molecule encodes a glyphosate-tolerant 5-enolipyruvil shikimate 3-phosphate synthase (EPSPS).

20. The method according to claim 17, further characterized in that said herbicide is glyphosate. 5 21. The method according to claim 20, further characterized in that said glyphosate is applied simultaneously with development in an effective amount of approximately 0.125 pounds of acid equivalent per acre and approximately 8 pounds of acid equivalent per acre.

22. A hybrid seed produced by the method as claimed in claim 17, the hybrid seed being characterized in that it comprises said recombinant DNA molecule.