| Vitis is one of the fruit trees that are widely cultivated in the world and have important economic value.Grape is also one of the main fruit trees in my country with a large cultivation area and high economic benefits,but its cold resistance is poor,and it needs to be buried in the soil to prevent cold in winter,which severely restricts the development of my country’s grape industry.Therefore,cultivating new grape germplasm that can adapt to the cold climate is one of the important tasks of our country’s breeders.In this experiment,the dormant buds of three F1 generation populations of grapes were used as test materials.Differential thermal analysis was used to identify the cold resistance of the dormant buds of grapes,combined with the genetic map constructed in the early stage of the laboratory for QTL mapping,and combined with transcriptome sequencing data Screening of candidate genes for grape cold resistance is expected to provide a theoretical reference for future grape cold resistance breeding.The main results are as follows:1.This test is based on the three grape combinations of,’Red Globe’×’Shuangyou’(RS),’Chardonnay’×’Beibinghong’(CB),’Cabernet Sauvignon’×’Zuoyouhong’(CZ)The dormant buds of the F1 generation population of European hybrid grapes were used as test materials.Differential thermal analysis was used to identify the cold resistance of the dormant buds of grapes,and the genetic information was analyzed.The cold resistance traits show the characteristics of quantitative inheritance,and a certain proportion of ultra-high cold resistance individual plants have been isolated in each population.2.In this experiment,QTL mapping was performed based on the cold hardiness phenotype data of grape dormant buds in 2019 and 2020,combined with the genetic map constructed in the early stage of the laboratory,and a total of 14 QTL loci that may be related to grape cold hardiness were located.LG2,LG3,LG4,LG6,LG9,LG14,LG15,LG16,LG17 on chromosomes.A stable QTL locus in different years was found,which is located on chromosome LG2 and chromosome LG3.3.In this experiment,transcriptome sequencing was performed on the dormant buds of parents at different stages under natural overwintering conditions,and a total of 15 candidate genes that may be related to grape cold resistance were screened,and q RT-PCR fluorescence quantitative verification was performed on them,of which 12 One cold-tolerance candidate gene was completely consistent with the transcriptome sequencing results,and the other three cold-tolerance candidate genes were basically consistent with the transcriptome sequencing results.4.In this experiment,transcriptome sequencing was performed on the dormant buds of parents at different stages under natural overwintering conditions,and a total of 15 candidate genes that may be related to grape cold resistance were screened,and q RT-PCR fluorescence quantitative verification was performed on them,of which 12 The cold-tolerance candidate genes are consistent with the results of transcriptome sequencing.5.This experiment cloned Vitvi02g00774(calmodulin),Vitvi02g00919(E3ubiquitin ligase),and Vitvi09g00044(CBF1).After sequence comparison and analysis,it was found that different parents VVitvi02g00919,Vitvi14g01394 and There is no difference in the CDS sequence of Vitvi09g00044,and the CDS sequence of Vitvi02g00774 is different in different parents.6.According to the allelic variation from C to T in the 148 th nucleotide of the Vv CML gene,this experiment developed the grape cold tolerance molecular marker Vach1,which was verified in 8 different grape varieties and 8 grape hybrid F1 generations. |
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