Map-based Cloning And Functional Analysing Of CUL1 Gene Involved In Leaf Rolling Form In Rice

Rice is one of the main food crops in the world,which plays an important role in ensuring the world food security.Leaf is the main place for photosynthesis.Therefore,it is of great theoretical significance and practical value to clone genes related to leaf morphological development and study their molecular mechanism for improving photosynthetic efficiency and rice yield.1.In this study,the curly leaf 1-1(cul1-1)mutant of cul1-1 was obtained by EMS mutagenesis with shennong9816 as the background material.In this study,cul1-1 was used as the test material to observe and analyze the leaf rolling phenotype,important agronomic characters and yield in detail.Cul1 gene was located and cloned by map based cloning.Using DNA recombination technology,the plant expression vector was constructed for complementary analysis.The main results of this study are as follows: using EMS mutagenesis,cul1-1 was obtained.The rolled leaf index of cul1-1 mutant was significantly higher than that of WT,and its heading stage was higher than that of tillering stage and seedling stage.2.Compared with the wild type,the fresh and dry weight of leaves,stems and ears of cul1-1 mutant decreased significantly.The ear length of cul1-1 had no significant change,but the plant height,tiller number and effective ear number were significantly lower than that of wild type.3.The theoretical yield per plant of cul1-1 mutant was significantly lower than that of wild type.Further analysis showed that the yield of cul1-1 mutant decreased due to the decrease of effective panicle number and seed setting rate.The decrease of seed setting rate of cul1-1mutant was mainly due to the increase of empty grain number,but there was no difference in total grain number.4.F2 mapping population was created by crossing cul1-1 mutant with habataki(HB).Genetic analysis showed that cul1-1 was a recessive mutation controlled by a single gene.5.The cul1 gene was cloned by map based cloning,and the mutation of cul1-1 was initially determined to be between the molecular markers B 7 and B 12 by Bulked aggregated analysis(BSA).Then,we expanded the population and developed new molecular markers.The mutation site of cul1-1 was located between SN 7 and SN 11.There are 32 open readingboxes.Using PCR and sequencing techniques,G to a base mutations were found in the 23 rd open reading frame,resulting in the transformation of amino acids from tryptophan to termination codon.6.The plant expression vector of complementary analysis was constructed.Using DNA recombination technology,UBI / 35S:CUL1-GFP plant expression vector was constructed.The method of Agrobacterium mediated genetic transformation of Rice Mature embryo was used to transfer cul1-1 mutant to obtain transgenic plants for complementary analysis.