中国水稻科学 ›› 2021, Vol. 35 ›› Issue (3): 238-248.DOI: 10.16819/j.1001-7216.2021.0604
刘维, 陆展华, 卢东柏, 王晓飞, 王石光, 薛皦, 何秀英*()
收稿日期:
2020-06-10
修回日期:
2020-08-06
出版日期:
2021-05-10
发布日期:
2021-05-10
通讯作者:
何秀英
基金资助:
Wei LIU, Zhanhua LU, Dongbai LU, Xiaofei WANG, Shiguang WANG, Jia XUE, Xiuying HE*()
Received:
2020-06-10
Revised:
2020-08-06
Online:
2021-05-10
Published:
2021-05-10
Contact:
Xiuying HE
摘要:
【目的】通过对水稻小穗簇生基因的定位与候选基因分析,为进一步克隆幼穗发育过程中缩短枝梗长度造成小穗簇生的功能基因奠定基础。【方法】以航天诱变育种技术处理常规优质稻品种粤农丝苗获得一个稳定遗传的小穗簇生突变体cl6为试验材料,与粤金银占构建F1、F2作图群体,对簇生基因进行定位,结合转录组测序对候选基因进行预测与评价。【结果】遗传分析结果表明,突变体cl6的簇生性状由非完全显性单基因控制,混合分组分析法将该基因定位于第6染色体上约416 kb的区间。进一步通过表达谱分析、转录组测序技术、基因序列差异性分析和qRT-PCR验证等筛选出OsFBK16为最佳候选基因,其5′-UTR区域发生9个碱基的插入突变,暗示该基因可能通过二级结构改变调控转录或翻译水平,参与水稻幼穗发育过程中枝梗的分化。【结论】本研究结果为解析水稻二次枝梗和小穗梗缩短机制奠定一定理论基础。
刘维, 陆展华, 卢东柏, 王晓飞, 王石光, 薛皦, 何秀英. 水稻小穗簇生基因OsCL6的定位及候选基因分析[J]. 中国水稻科学, 2021, 35(3): 238-248.
Wei LIU, Zhanhua LU, Dongbai LU, Xiaofei WANG, Shiguang WANG, Jia XUE, Xiuying HE. Location and Candidate Gene Analysis of Rice Clustered Spikelets Gene OsCL6[J]. Chinese Journal OF Rice Science, 2021, 35(3): 238-248.
基因登录号Gene ID | 基因注释Gene annotation | 备注Remark |
---|---|---|
LOC_Os06g39260 | 溶质载体家族35成员B1 Solute carrier family 35 member B1, putative, expressed | OsUGT41[ |
LOC_Os06g39344 | 烯醇辅酶A水合酶/异构酶家族基因Enoyl-CoA hydratase/isomerase family protein, putative | |
LOC_Os06g39370 | OsFBK16-含F盒和Kelch结构域的蛋白质OsFBK16-F-box domain and kelch repeat containing protein | |
LOC_Os06g39480 | 含SPOC结构域的蛋白质SPOC domain containing protein, putative, expressed | |
LOC_Os06g39600 | 蛋白磷酸酶2C蛋白Protein phosphatase 2C, putative, expressed | |
LOC_Os06g39640 | PINHEAD, putative, expressed | OsPNH1[ |
LOC_Os06g39680 | 转座子蛋白 Transposon protein, putative, unclassified | |
LOC_Os06g39750 | 3-酮酰基-coA合酶3-ketoacyl-CoA synthase, putative, expressed | WSL1[ |
LOC_Os06g39760 | WD结构域,含G-β重复结构域的蛋白质WD domain, G-beta repeat domain containing protein | |
LOC_Os06g39870 | 26S蛋白酶调节亚基8 26S protease regulatory subunit 8, putative | |
LOC_Os06g39880 | 细胞色素P450 72A1 Cytochrome P450 72A1, putative, expressed | CYP734A4[ |
表1 幼穗发育时期高表达的11个基因
Table 1 Eleven highly expressed genes in young panicle development.
基因登录号Gene ID | 基因注释Gene annotation | 备注Remark |
---|---|---|
LOC_Os06g39260 | 溶质载体家族35成员B1 Solute carrier family 35 member B1, putative, expressed | OsUGT41[ |
LOC_Os06g39344 | 烯醇辅酶A水合酶/异构酶家族基因Enoyl-CoA hydratase/isomerase family protein, putative | |
LOC_Os06g39370 | OsFBK16-含F盒和Kelch结构域的蛋白质OsFBK16-F-box domain and kelch repeat containing protein | |
LOC_Os06g39480 | 含SPOC结构域的蛋白质SPOC domain containing protein, putative, expressed | |
LOC_Os06g39600 | 蛋白磷酸酶2C蛋白Protein phosphatase 2C, putative, expressed | |
LOC_Os06g39640 | PINHEAD, putative, expressed | OsPNH1[ |
LOC_Os06g39680 | 转座子蛋白 Transposon protein, putative, unclassified | |
LOC_Os06g39750 | 3-酮酰基-coA合酶3-ketoacyl-CoA synthase, putative, expressed | WSL1[ |
LOC_Os06g39760 | WD结构域,含G-β重复结构域的蛋白质WD domain, G-beta repeat domain containing protein | |
LOC_Os06g39870 | 26S蛋白酶调节亚基8 26S protease regulatory subunit 8, putative | |
LOC_Os06g39880 | 细胞色素P450 72A1 Cytochrome P450 72A1, putative, expressed | CYP734A4[ |
图3 候选基因在幼穗中优势表达的模式1-萌发后的胚和胚根; 2-浸泡72 h后的种子;3-二蘖苗的根;4-二蘖苗的地上部;5-3叶期的叶和根;6-幼穗分化Ⅲ期的叶片;7-幼穗长达4~5 cm时的叶片;8-抽穗前5 d的剑叶;9-抽穗前14 d的剑叶;10-幼穗分化Ⅲ期的茎鞘;11-幼穗长达4~5 cm时的茎鞘;12-抽穗前5 d的茎鞘;13-抽穗前1 d的颖壳;14-抽穗期的茎;15-处于分化Ⅲ期的幼穗;16-处于分化Ⅳ期的幼穗;17-处于分化V期的幼穗;18-4~5 cm长幼穗;19-抽穗期的穗;20-开花前1 d的雄蕊;21-授粉3 d后的小穗;22-授粉7 d后的胚乳;23-授粉14 d后的胚乳;24-授粉21 d后的胚乳。
Fig. 3. Patterns of dominant expression of candidate genes in young panicles. 1, Embryo and radicle after germination; 2, Seed at 72 h after imbibition; 3, Root of seedling with 2 tillers; 4, Shoot of seedling with 2 tillers; 5, Leaf and root at 3-leaf stage; 6, Leaf at stage Ⅲ of young panicle differentiation; 7, Leaf, at 4-5-cm young panicle stage; 8, Flag leaf, 5 days before heading; 9, Flag leaf, 14 days before heading; 10, Sheath, at stage Ⅲ of panicle differentiation; 11, Sheath, at 4-5-cm young panicle stage; 12, Stem, 5 days before heading; 13, Hull, one day before flowering; 14, Stem, heading stage; 15, Young panicle at stage Ⅲ of panicle differentiation; 16, Young panicle stage Ⅳ of young panicle differentiation; 17, Young panicle at stage V of panicle differentiation; 18, Panicle at 4-5-cm young panicle stage; 19, Panicle, heading stage; 20, Stamen, one day before flowering; 21, Spikelet, 3 days after pollination; 22, Endosperm, 7 days after pollination; 23, Endosperm, 14 days after pollination; 24, Endosperm, 21 days after pollination.
图4 候选基因OsFKB16验证 A-采用qRT-PCR验证OsFKB16基因的表达差异; B-OsFKB16的启动子测序结果; C-OsFKB16的启动子共分离验证。WT-粤农丝苗。
Fig. 4. Candidate gene OsFKB16 validation. A, qRT-PCR analysis expression difference of candidate gene OsFKB16; B, Results of OsFKB16 promoter sequencing; C, OsFKB16 promoter co-separation validation. WT, Yuenongsimiao.
图5 野生型与突变体转录组测序部分分析结果 A-差异表达基因聚类分析,log2(FPKM+1); B-差异表达基因KEGG通路富集散点图;C-GA合成代谢和信号途径相关基因的表达差异。
Fig. 5. Partial analysis results of transcriptome sequencing of the wild type and its mutant. A, Cluster map of differentially expressed genes, log2(FPKM+1); B, Enrichment scatter map of differentially expressed gene KEGG pathway; C, Expression difference of some genes related to GA anabolism and signaling pathway.
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