| Rice is not only a very important food crop,but also an important model crop to study genetic functions.Rice yield is constitutes by four factors: panicle number per unit area,grain number per panicle,seed setting rate and 1000-grain weight.As the major pool of rice,the development of panicle plays a vital role in the yield and quality of rice.Some related genes were screened with the chip related to panicle development at the beginning,and lots of corresponding mutants were obtained to explore functions of these genes above via CRISPR/Cas9 technology.We isolated a gene named OsD1 and obtained knock-out mutants with CRISPR/Cas9,which was used for phenotypic observation and functional analysis.The findings are as follows:1?A rice gene OsD1 was analyzed with bioinformatics.The genomic sequence length of it is 2623 bp with 3 exons and 2 introns and CDS length of it is 1038 bp encoding a conserved protein sequence which contains 345 amino acids.OsD1 belongs to the R2R3-MYB subfamily due to it encodes a transcription factor containing two MYB functional domains.2?The analysis of OsD1 expression profile shows that the expression of this gene increases with the continuous development of young panicle,and expression signal detected in stamen,pistil,palea and lemma are in different degrees.3 ? The CRISPR/Cas9 site-directed editing vector of OsD1 was constructed and successfully transformed into rice callus.A batch of mutants were identified and there were several biallelic homozygous mutants and biallelic heterozygous mutations among them.4?The phenotype identification of the biallelic homozygous mutant revealed that from the heading stage to the mature stage of rice,there were several obvious malformation development in palea and lemma,and palea developed into lemma.The size of glume increased.Moreover,the anther at the heading stage showed the presence of several microspore with starch of low plumpness by being examined with I2-KI on the microscopy. |
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