DELLA gene families of peanut as well as encoding genes and applications of DELLA gene families

A technique for peanuts and encoding, which is applied in the field of molecular biology and can solve problems such as the backwardness of peanut molecular biology research

CN104558132AActive Publication Date: 2015-04-29BIOTECH RES CENT SHANDONG ACADEMY OF AGRI SCI
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Patent Information

Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Publication Date
2015-04-29

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Abstract

The invention relates to DELLA gene families of a peanut as well as encoding genes and application of the DELLA gene families and discloses an AhDELLA3 protein of the peanut and an AhDELLA4 protein of the peanut, wherein the amino acid sequence of the AhDELLA3 protein is as shown in SEQ ID NO.5; and the amino acid sequence of the AhDELLA4 protein is as shown in SEQ ID NO.7. The invention firstly finds the DELLA protein gene families AhDELLA3 and AhDELLA4 of the peanut and encoding genes thereof, finds important applications of AhDELLA3 and AhDELLA4 genes to the pod development process of the peanut through analyzing the expression changes of the AhDELLA3 and AhDELLA4 genes in different tissues of the peanut, provides the important theoretical basis for the molecular mechanism of the DELLA gene families on the aspect of controlling the growth and development of the peanut, has important guiding significance on the aspect of cultivating new species of peanuts, and lays the foundation for innovating novel germplasms for crops by using a transgenic technology.
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Description

technical field

[0001] The invention relates to the peanut DELLA gene family and its encoding gene and application, in particular to the cloning, expression analysis, function identification and application of the peanut DELLA gene family, and belongs to the technical field of molecular biology. Background technique

[0002] Gibberellins (GAs for short), as a plant hormone, play a very important role in the regulation of plant growth and development. For example: promoting seed germination, promoting stem elongation and leaf growth, inducing flowering and seed and fruit growth and other physiological functions.

[0003] DELLA proteins are inhibitors in the gibberellin signaling pathway (see Fleet et al. Current Opinion in Plant Biology, 877-85, 2005). The gibberellin signaling pathway is mainly realized by releasing the inhibitory effect of DELLA protein. After receiving the GA signal, the receptor GID1 forms a GA-GID1-DELLA complex with the DELLA protein, which is then re...

Examples

Embodiment 1

[0049] Example 1: Cloning of Peanut AhDELLA Gene Family Gene and Full-length Gibberellin Receptor AhGID1 Gene

[0050] Using the nucleotide sequence of soybean DELLA protein with high homology to peanut, the homologous comparison was used to search the early transcriptome database of peanut needle and pod development, and a total of four groups of Unigenes related to DELLA were found, and each group came from one gene. Among them, the Unigene from AhDELLA2 contains a start codon and is deleted at the 3' end; the Unigenes from AhDELLA1, AhDELLA3, and AhDELLA4 are all deleted at the 5' end and 3' end. The same method was used to search the transcriptome database in the early development of peanut needles and pods, and the full-length sequence of the AhGID1 gene was obtained by splicing short fragments.

[0051] Nested PCR was amplified by 5'RACE and 3'RACE, and the full-length open reading frames of the DELLA gene family AhDELLA1, AhDELLA2, AhDELLA3 and AhDELLA44 genes were fina...

Embodiment 2

[0155] Example 2: Homology analysis of peanut DELLA protein

[0156] The amino acid sequences of the peanut DELLA protein gene family are shown in SEQ ID NO.1, SEQ ID NO.3, SEQ ID NO.5, and SEQ ID NO.7.

[0157] Search the DELLA protein of the following species on the NCBI website: Arabidopsis thaliana, wheat, soybean, barley, rice, castor, pea, and use the DNAMAN software to compare the amino acid sequence homology ( figure 1 ). The results show that AhDELLA1 and AhDELLA2 are divided into two branches, and the amino acid sequence identities with DELLA proteins of other species are about 70% and 65%, respectively; while the homology between AhDELLA3 and AhDELLA4 is very high, reaching 97%, but Homology to other species is low. It shows that AhDELLA3 and AhDELLA4 may have similar functions, but unlike AhDELLA1 and AhDELLA2, AhDELLA3 and AhDELLA4 may have unique functions in peanut.

Embodiment 3

[0158] Example 3: Analysis of the expression pattern of the DELLA gene family in different tissues of peanut

[0159] 3.1 Extraction of RNA from plants:

[0160] With the 1.1 step in the embodiment 1, extract respectively peanut juvenile stem (15 days, JS), adult stem (40 days, AS), juvenile leaf (15 days, JL), adult leaf (40 days, AL), root ( JR), flowers (F), fruit needles not buried in soil (G0), fruit needles buried in soil for 3 days (G3), fruit needles buried in soil for 9 days (G9), immature seeds (S) and other tissues in different periods. RNA.

[0161] 3.2 Acquisition of cDNA from different tissues of peanut:

[0162] Same as 1.11 step (1) in Example 1.

[0163] 3.3 Using fluorescent quantitative PCR technology to analyze the expression pattern of DELLA gene family genes:

[0164] With AhActin as the system internal reference, the primer sequence is:

[0165] AhActinqF:5'-GTCATCGTCATCCTCTTCTC-3'(SEQ ID NO.35);

[0166] AhActinqR: 5'-CATTCCTGTTCCATTGTCAC-3' (SEQ ...