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Use of rice three-flower spikelet gene LF1 in increase of yield of gramineous food crops

A technology for food crops and rice, applied in the field of molecular biology, can solve the problems of lack of application value, not and other problems, and achieve the effect of improving rice yield and increasing the number of grains per ear

Active Publication Date: 2017-09-08
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0014] Although the studies of G1, OsMADS34 / PAP2, EG1, NSG, ASP1, and OsMADS1 / LHS1 genes provided some support for the hypothesis of ―3 flower spikelets, suggesting that glume may be the degenerated lemma of lateral florets, but in these In mutants or transgenic materials, glumes only "recover" into lemma-shaped organs or elongate and become larger, but do not "recover" into lateral florets with inner flower organs such as pistils, which is not a true lemma. ―3 flower spikelets”, lack of application value

Method used

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  • Use of rice three-flower spikelet gene LF1 in increase of yield of gramineous food crops
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  • Use of rice three-flower spikelet gene LF1 in increase of yield of gramineous food crops

Examples

Experimental program
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Effect test

Embodiment 1

[0046] Example 1 Obtaining and Phenotype Analysis of lf1 Mutant

[0047] Ethyl methanesulfonate (EMS) was used to mutate the self-fertile Jiahui Jinhui 10. In the offspring mutant population, a genetically stable gain-of-function (dominant) mutant of rice -3 flower spikelet was found. Multi-generation self-crossing, genetic stability, named lateral floret 1 (lf1).

[0048] At the heading stage of rice, lf1 and wild-type spikelets were taken for morphological and histological analysis. Morphological analysis: On the one hand, fresh spikelets were observed, dissected and photographed under a Nikon SM1500 stereomicroscope; on the other hand, fresh spikelets were observed under a scanning electron microscope at -40° using a Hitachi SU3500 scanning electron microscope. Histological analysis: fresh spikelets were fixed overnight at 4° in FAA fixative solution (50% ethanol, 0.9M glacial acetic acid and 3.7% formaldehyde), dehydrated through ethanol gradient, xylene replacement, and ...

Embodiment 2

[0053] Example 2 Genetic analysis, gene mapping and map-based cloning of LF1 gene

[0054] The lf1 mutant was crossed with Xida 1A (provided by the Rice Research Institute of Southwest University). A total of 404 F strains with the same phenotype as the lf1 mutant were obtained. 2 Plants were used for genetic analysis and gene mapping of LF1. f 2 The segregation population showed that the segregation ratio between mutant and wild-type plants was 3:1, suggesting that the phenotype of the lf1 mutant was controlled by a pair of dominant nuclear genes.

[0055]Preliminary positioning: BSA (mixed cluster analysis) method was used. Select 480 pairs of SSR primers evenly distributed on the 12 chromosomes of rice Gramene, the Rice Genomic Research Program website, and according to the insertion / deletion fragment difference between Xida 1A and Jinhui 10, SNP (single nucleotide Polymorphisms) self-developed in / del and dCAPs marker primers (ZIn23F / R, ZIn29F / R, ZIn27F / R, Zin67F / R and ...

Embodiment 3

[0061] Example 3 Analysis of the expression pattern of the LF1 gene

[0062] Using real-time quantitative PCR (the sequences of primers qLF1-F and qLF1-R are shown in Table 1, Actin is used as an internal standard, and the sequences of primers Actin-F and Actin-R are shown in Table 1) to detect the presence of LF1 gene in the young panicle and each round flower organ The results showed that LF1 was expressed in young panicles of different periods, especially in young panicles below 2 cm, which were significantly higher than those in other periods. At the heading stage, LF1 was expressed in the glume and the four round flower organs ( Figure 5 a).

[0063] In situ hybridization was used to further analyze the expression pattern of LF1 gene in spikelets (see Table 1 for the sequences of primers LF1-F and LF1SP6-R). At the Sp4 stage, LF1 was abundantly expressed in the terminal florets of wild-type and lf1 mutant spikelets ( Figure 5 b1, b5). From Sp5 to Sp6, LF1 was expres...

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Abstract

The invention belongs to the technical field of molecular biology and particularly relates to use of a rice three-flower spikelet gene LF1 in the increase of the yield of gramineous food crops. The invention further provides the use of the rice three-flower spikelet gene LF1 in the increase of the yield of the gramineous food crops and a preparation for increasing the yield of the gramineous food crops. According to the use, the gene LF1 is separated and causes ectopic expression due to the mutation of a microRNA165 / 166 regulatory site, so that two ''lateral florets'' in a spikelet are restored, and the number of the florets of the spikelet of an lf1 mutant reaches 3 (named as a ''three-flower spikelet'') together with an original acrogenous fertile floret. A new breeding way for increasing the kernel number per spike is provided and is of a very important significance to the further increase of the yield of the rice. A new choice is provided for the increase of the yield of the gramineous food crops.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to the use of rice 3-flower spikelet gene LF1 in increasing the yield of gramineous grain crops. Background technique [0002] The number of grains per panicle is one of the important factors that constitute the yield of gramineous grain crops such as rice, and the morphological traits of panicles (inflorescences), such as the number of branches at all levels and the density of spikelets on them, will affect the number of grains per panicle. The formation of grain number, the currently cloned genes related to the number of grains per panicle also mainly affect the panicle morphological traits. In fact, there is another important factor affecting the formation of the number of grains per panicle—the number of florets in spikelets. This phenomenon has not attracted people’s attention, because the number of florets in a spikelet of normal rice is constant—only includes A fer...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00
Inventor 李云峰张婷何光华桑贤春凌英华杨正林
Owner SOUTHWEST UNIVERSITY
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