Development and application of a molecular marker for low glutelin content gene in rice lenshuinuo | 水稻品种冷水糯的低谷蛋白性状连锁标记的开发与应用

Chinese. 摘　　要：我国肾脏病患者和糖尿病肾病患者呈不断增加的趋势,筛选和培育这一特殊人群可以食用的谷蛋白含量低的水稻品种具有重要意义。在前期研究中获得一个低谷蛋白隐性突变体冷水糯,该突变体籽粒的谷蛋白具有57kD前体聚增（57H）伴有成熟亚基减少。由于glup3基因与57H前体裂解相关,本研究对冷水糯的glup3测序、比对后,发现冷水糯的glup3基因的编码区第824个碱基由G突变为A,导致翻译产物第275个氨基酸由丝氨酸变为天冬酰胺。根据突变位点两侧的序列设计了CAPS标记DG-2,该标记能扩增出173bp的条带,运用TscAI酶对该片段进行酶切,来自于冷水糯的片段不能被该酶切开,来自于9311的片段可被切成105bp和68bp的两条片段,而来自于9311/冷水糯的杂交1代PCR产物经酶切后,出现双亲的带型。该标记对9311/冷水糯的高代品系（F7代）的100个家系进行鉴定,结合SDS-PAGE蛋白电泳结果,证实该标记可以准确地区分具有57H前体聚增的低谷蛋白水稻材料。同时,该标记可以区分glup3基因的杂合型,可以用于分子标记辅助选择育种。

English. Lenshuinuo, a low-glutelin-content （LGC） rice mutant, was selected to develop molecular marker for LGC assistant selection. SDS-PAGE electrophoresis showed that Lenshuinuo belonged to 57H LGC rice variety, so, its glup3 gene was screened out to study the gene difference with ordinary rice variety 9311 because glup3 gene was relative to 57H incision. The results showed that the 824th base in coding region of glup3 gene of Lenshuinuo changed from G to A, which leaded to the 275th amino acid variation from serine to asparagine comparing with 9311. A CAPS marker, DG-2, were designed based on the sequence flanked the mutation site and its PCR product （173 bp） from Lenshuinuo couldn＇t be cut into two parts （105 bp and 68 bp） by TscAI enzyme while its PCR product （173 bp） from 9311 reverse. F1 generation and 100 advanced ancestries （F7 generation） from 9311/Lenshuinuo were identified using DG-2 markers and SDS-PAGE electrophoresis. the result showed that the glutelin contents from endosperm of the ancestries with PCR product not be cut were low; Enzyme-cutting PCR product from F1 generation exhibited 3 bands （173 bp, 105 bp and 68 bp）, So this marker can be used to differentiate ancestry heterozygosity and the molecular assistant selection （MAS） for low-glutelin-content rice variety from Lenshuinuo hybrid generation.