Gene for regulating and controlling maturing rate and application thereof
A technology of seed setting rate and genetics, applied in the fields of biotechnology and botany
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Embodiment approach
[0151] As an embodiment of the present invention, the PSSR1 gene (without transposon) is cloned into an appropriate vector by conventional methods, and the recombinant vector with the foreign gene is introduced into plant tissues or organs , so that the plant expresses the PSSR1 gene. A plant overexpressing the PSSR1 gene can be obtained by regenerating the plant tissue or organ into a plant.
[0152] Alternatively, the transposon (insert sequence) in the low-activity PSSR1 gene can be knocked out by means known to those in the art, and / or the transposon can be edited using gene editing techniques such as ZFN, TALEN or CRISPR / Cas9, etc. The vector with knockout, editing or transposition function is introduced into plant tissues or organs to restore PSSR1 gene expression or activity. The plant tissue or organ can be regenerated into a plant to obtain a plant that restores the expression of the PSSR1 gene. The structure of such knockout, editing or transposition vectors, such ...
Embodiment 1
[0191] Example 1, WinQTL analysis of seed setting rate traits
[0192] 250 BIL and CSSL lines were investigated for seed setting rate traits, and WinQTL analysis was performed to calculate QTL loci related to seed setting rate. Two major QTL loci were found, which were located on chromosome 1 (43.9-44.3Mb) and chromosome 9, of which the QTL locus on chromosome 9 was a known gene ( figure 1 ).
Embodiment 2
[0193] Example 2, map-based cloning
[0194] Through 14 pairs of Indel and SNP molecular markers at the end of chromosome 1, the unknown seed setting rate-related genes were initially located in the 49Kb range between 44.042cM and 44.091cM at the end of the long arm of chromosome. There are a total of 7 predicted genes in this 49Kb range ( figure 2 ).
[0195] The expression levels of 7 predicted genes in the panicle of GLA4 and NIL-PSSR1 plants were analyzed by q-PCR, and it was found that the expression level of orf4 was significantly different between GLA4 and NIL-PSSR1 plants, and orf4 was in NIL-PSSR1. hardly express. In addition, the expression levels of orf6 and orf7 in the panicle of GLA4 and NIL-PSSR1 were extremely low, and there was no significant difference; although the expression levels of orf1, orf2, orf3 and orf5 were all slightly higher in NIL-PSSR1 than in GLA4, the No significant difference (P>0.01) ( image 3 ). Therefore, we selected orf4 as a candid...
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