大豆荚皮和种皮高表达的三个蔗糖合酶基因的表达与功能分析  

Expression and functional analysis of three sucrose synthase genes highly expressed in pod coats and seed coats of soybean

作　　者：黄俊铭 傅旭军[3] 寿惠霞 HUANG Junming;FU Xujun;SHOU Huixia(Advanced Seed Institute,College of Agriculture and Biotechnology,Zhejiang University,Hangzhou 310058,China;Seed Engineering and Industrialization Group,Hainan Institute of Zhejiang University,Sanya,Hainan 572000,China;institute of Crop and Nuclear Technology Utilization,Zhejiang Academy of Agricultural Sciences,Hangzhou 310021,China)

机构地区：[1]浙江大学农业与生物技术学院现代种业研究所,杭州310058 [2]浙江大学海南研究院种子工程与产业化团队,海南三亚572000 [3]浙江省农业科学院作物与核技术利用研究所,杭州310021

出　　处：《植物生理学报》2024年第7期1119-1131,共13页Plant Physiology Journal

基　　金：国家重点研发计划(2021YFF1001204)。

摘　　要：蔗糖是高等植物光合产物的主要运输形式,蔗糖合酶(sucrose synthase,SUS)是一类糖基转移酶,在库组织同化产物的代谢中发挥关键作用,影响作物产量和品质。本研究前期通过RNA-Seq数据分析,发现大豆SUS5亚家族基因——GmSUS5a、GmSUS5b、GmSUS5c在豆荚和种子中高表达,但它们的时空表达模式和功能尚有待解析。生物信息学分析表明,3个GmSUS5s蛋白相似度在82.0%以上,编码840~845个氨基酸,定位于细胞质中。对不同发育阶段荚皮和种子取样分析表明,三基因在花后7周鼓粒期的荚皮与种皮中表达最高,以Gm SUS5a表达量最高。对基因编辑获得的GmSUS5单、双和三突变体进行了表型观察,发现GmSUS5突变体因豆荚和种胚糖供应不足,造成坐果困难花期延长。本研究明确了大豆GmSUS5s参与种子蔗糖供应,从而影响大豆种子的发育。研究结果为进一步研究大豆蔗糖合酶基因GmSUS5s的功能和作用机制奠定了基础。Sucrose is the main transport form of photosynthetic product in higher plants.Sucrose synthase(sUs)is a glycosyl transferase enzyme that plays a key role in sugar metabolism,primarily in sink tissues,therefore affects crop yield and quality.RNA-Seq data in our previous study showed that the soybean SUS5 subfamily genes,GmSUS5a,GmSUS5b and GmSUS5c were highly expressed in pods and seeds.However,their spatio-temporal expression patterns and functions have yet to be analyzed.Bioinformatics analysis showed that the GmsUS5s proteins share 82.0%and greater sequence similarity,encoding 840-845 ami-no acids and localized in cytoplasm.RT-qPCR analysis of pod coats and seeds at different developmental stages showed that the expression of the GmSUS5 genes reached at the highest level in pods and seed coats at seed filling stage,seven weeks after flowering,and the expression level of GmsUS5a was the highest.The single,double and triple mutants of GmSUS5 genes obtained by CRISPR/Cas9 gene editing showed prolonged flowering period and inefficiency in pod-setting,which may be due to the insufficient sugar supply to pods and embryos.This study confirmed that GmSUS5s are involved in seed sucrose supply,thereby affecting soybean seed development.This study lays a foundation for further study of the function and mechanism of soybean sucrose synthases genes GmSUS5.

关 键 词：大豆(Glycine max) 蔗糖合酶 GmSUS5 基因表达 基因编辑 

分 类 号：S565.1[农业科学—作物学]