南瓜淀粉组分高效检测体系建立及应用  

Establishment and Application of Efficient Detection System for Starch Components in Pumpkin

作　　者：宋慧 安学君 黄芸萍 沈登锋 吕燕 陈国 陈山乔 张香琴 SONG Hui;AN Xuejun;HUANG Yunping;SHEN Dengfeng;LV Yan;CHEN Guo;CHEN Shanqiao;ZHANG Xiangqin(Ningbo Academy of Agricultural Sciences,Ningbo,Zhejiang 315040)

机构地区：[1]宁波市农业科学研究院,宁波315040

出　　处：《中国农学通报》2024年第5期135-144,共10页Chinese Agricultural Science Bulletin

基　　金：宁波市公益类科技计划项目“类胡萝卜素与淀粉双高粉用加工型南瓜品种辅助选育与示范”(2021S016);国家自然科学基金“果糖激酶对瓜类异属间嫁接亲和特性的指示作用和调控机理”(31572142)。

摘　　要：旨在建立高效的南瓜淀粉组分检测技术,以期提高定向育种和合理开发南瓜新品种的效率。根据双波长比色原理,利用提纯的南瓜直链淀粉和支链淀粉,通过等吸收点作图法,确定南瓜淀粉组分测定波长和参比波长。通过单因素比较试验,确定双波长测定南瓜淀粉组分的200μL反应体系。最后比较不同类型的南瓜淀粉组分含量,筛选极端材料。结果表明,直链淀粉的测定波长为616 nm、参比波长为472 nm,南瓜支链淀粉的测定波长为545 nm、参比波长为725 nm。建立南瓜直链淀粉和支链淀粉回归方程,直链淀粉浓度在0.0125~1 mg/mL、支链淀粉浓度在0.025~2 mg/mL范围内,线性关系良好(R2>0.9900)。96孔板200μL反应体系为待测液20μL、醋酸盐缓冲反应液(pH 3.5)90μL、12倍碘液稀释液90μL。供试23份南瓜材料的直链淀粉含量在0.52~26.68 mg/g果肉鲜重,支链淀粉含量在0.03~76.13 mg/g果肉鲜重,淀粉总量在0.55~101.61 mg/g果肉鲜重。以10和15 mg/g果肉鲜重、35和45 mg/g果肉鲜重、55和65 mg/g果肉鲜重,可以分别初步评价供试南瓜直链淀粉、支链淀粉和总淀粉含量低、中、高的情况。中印南瓜自交系比供试中国南瓜和印度南瓜的淀粉组分含量均高,最终筛选到7个中印南瓜自交系,用以淀粉定向育种潜力大。该试验获得的南瓜淀粉组分高效检测体系,对提高南瓜淀粉育种效率,行之有效。To establish the efficient determination of pumpkin starch components,which would be of great significance for directional breeding of special pumpkin starch varieties and rational development,the experiments were carried out.According to the dual-wavelength colorimetric principle,the determination wavelength and reference wavelength of pumpkin starch components were determined by using the purified pumpkin amylose and amylopectin and the equal absorption point diagram.The 200μL reaction system for the determination of pumpkin starch components with dual wavelengths was determined by single factor comparison test.Finally,the content of starch components in different types of pumpkin was compared.The extreme materials were screened.The results showed that the determination wavelength of pumpkin amylose was 616 nm,the reference wavelength was 472 nm,and the determination wavelength of pumpkin amylopectin was 545 nm and the reference wavelength was 725 nm.The regression equations of pumpkin amylose and amylopectin were established.The linear relationship was good(R2>0.9900)when the amylose concentration was in the range of 0.0125-1 mg/mL,and the amylopectin concentration was in the range of 0.025-2 mg/mL.The 200μL reaction system in a 96-well plate consisted of 20μL of sample,90μL of acetate buffered saline(pH 3.5),and 90μL of 12-fold iodine diluent.The tested 23 pumpkin materials had amylose content of 0.52-26.68 mg/g fresh flesh weight,amylopectin content of 0.03-76.13 mg/g fresh flesh weight and total starch content of 0.55-101.61 mg/g fresh flesh weight.The fresh flesh weight of 10 and 15 mg/g,the fresh flesh weight of 35 and 45 mg/g,and the fresh flesh weight of 55 and 65 mg/g could preliminarily evaluate the low,medium and high contents of amylose,amylopectin and total starch in tested pumpkin,respectively.The content of starch components of the inbred lines derived from inter-specific hybrids of Cucurbita moschata Duch.×Cucurbita maximam Duch.were higher than that of the tested C.moschata and C.maximam.

关 键 词：南瓜 直链淀粉 支链淀粉 淀粉含量 远缘杂交 

分 类 号：S642.1[农业科学—蔬菜学]