黄淮麦区部分小麦种质脂肪氧化酶活性分析及等位基因检测

doi:10.3864/j.issn.0578-1752.2015.02.01

摘要/Abstract

摘要： 【目的】小麦中的脂肪氧化酶（LOX）是影响小麦面粉颜色和其储藏特性的主要因素之一，对中国黄淮麦区的306份小麦种质资源进行脂肪氧化酶活性分析及等位基因检测，为小麦品质育种提供理论参考。【方法】利用紫外分光光度计和酶标仪测定参试样品的脂肪氧化酶（LOX）活性，并利用控制脂肪氧化酶活性的位于1AL上的QLpx.caas-1AL位点紧密连锁的分子标记Xwmc312以及位于4BS上的TaLOX-B1位点的功能标记LOX16和LOX18，采用特异引物的PCR扩增技术以及琼脂糖凝胶电泳和聚丙烯酰胺凝胶电泳分离技术进行参试材料的LOX基因型鉴定。【结果】表型测定结果表明，参试的黄淮麦区小麦品种（系）中脂肪氧化酶活性的平均值为65.73 AU·min^-1·g^-1，标准差为13.54，变幅为27.09—99.55 AU·min^-1·g^-1，变异系数为20.6%。参试材料中小于40 AU·min^-1·g^-1和大于90 AU·min^-1·g^-1的小麦品种（系）分别为7个和8个，可作为对当前主栽小麦品种进行脂肪氧化酶活性改良的重要资源材料。基因型鉴定结果表明，参试材料的QLpx.caas-1AL位点存在3种等位变异类型，Xwmc312₂₂₇、Xwmc312₂₃₅和Xwmc312₂₄₇所占比例分别为30.4%、18.9%和50.6%。参试材料的TaLOX-B1位点存在TaLOX-B1a和TaLOX-B1b两种变异类型，所占比例分别为 28.7%和71.2%。分析其基因型组合发现，参试材料中共有6种基因型组合类型，依次为TaLOX-B1a/Xwmc312₂₂₇、TaLOX-B1a/Xwmc312₂₃₅、TaLOX-B1a/Xwmc312₂₄₇、TaLOX-B1b/Xwmc312₂₂₇、TaLOX-B1b/Xwmc312₂₃₅和TaLOX-B1b/Xwmc312₂₄₇，所占比例分别为7.2%、9.5%、12.1%、23.2%、9.5%和38.6%。分析不同LOX基因型与脂肪氧化酶活性的关系表明，1AL位点上拥有Xwmc312₂₃₅基因型的小麦品种脂肪氧化酶活性显著高于其他两种基因型（P＜0.05），4BS位点拥有TaLOX-B1a基因型的小麦品种脂肪氧化酶活性显著高于拥有TaLOX-B1b基因型的小麦品种（P＜0.05）。进一步分析不同LOX基因型组合与脂肪氧化酶活性的关系表明，拥有TaLOX-B1a/Xwmc312₂₃₅基因型组合的小麦品种脂肪氧化酶活性（76.80 AU·min^-1·g^-1）显著高于其他5种基因型组合，而拥有TaLOX-B1b/Xwmc312₂₂₇基因型组合的小麦品种脂肪氧化酶活性显著低于其他5种基因型组合，仅为62.44 AU·min^-1·g^-1。【结论】中国黄淮麦区的小麦品种脂肪氧化酶活性绝大多数处于中间类型，拥有极低（小于40 AU·min^-1·g^-1）或极高（大于90 AU·min^-1·g^-1）脂肪氧化酶活性类型的小麦品种所占比例较低。黄淮麦区所发现的6种不同LOX基因型组合中，拥有TaLOX-B1a/Xwmc312₂₃₅基因型组合的小麦品种脂肪氧化酶活性相对较高（P＜0.05），而拥有TaLOX-B1b/Xwmc312₂₂₇基因型组合的小麦品种脂肪氧化酶活性相对较低（P＜0.05）。

关键词: 普通小麦, 面粉颜色, 脂肪氧化酶, 等位变异

Abstract: 【Objective】The objective of this study is to provide useful information for wheat breeding program, a total of 306 wheat cultivars or advanced lines from the Yellow and Huai valleys of China were used to identify LOX (Lipoxygenase) activity and allelic variations of LOX genes in this study.【Method】 Spectrophotometer and microplate were used to measure the LOX activity. The molecular marker Xwmc312 on chromosome 1AL and the functionalmarkers LOX16 and LOX18 on chromosome 4BSwere employed to identify the polymorphism of the LOX genes in Chinese wheat cultivars or advanced lines surveyed by combinations of PCR (Polymerase Chain Reaction) amplification and agarose and electrophoresis gels. 【Result】 The phenotyping results showed that the averaged LOX activity was 65.73 AU·min^-1·g^-1 and the standard deviation (SD) was 13.54 in the wheat cultivars surveyed from the Yellow and Huai valleys, and it ranged from 27.09 to 99.55 AU·min^-1·g^-1 and its coefficient of variation (CV) was 20.6%. In the wheat cultivars surveyed from the Yellow and Huai valleys, LOX activities of 7 wheat cultivars were less than 40 AU·min^-1·g^-1 and LOX activities of 8 wheat cultivars exceeded 90 AU·min^-1·g^-1. These cultivars could provide very important germplasms for improvement of LOX activities of currently popular wheat in wheat breeding program. Genotyping results indicated that three alleles Xwmc312₂₂₇, Xwmc312₂₃₅ and Xwmc312₂₄₇were found at the QLpx.caas-1AL locus in these wheat varieties with 30.4%, 19.0% and 50.6%, respectively. At TaLOX-B1 locus, two alleles TaLOX-B1a and TaLOX-B1b were identified by the functionalmarkers LOX16 and LOX18 with 28.8% and 71.3%, respectively, in the bread wheat cultivars surveyed. Six allele combinations of LOX genes, i.e., TaLOX-B1a/Xwmc312₂₂₇, TaLOX-B1a/Xwmc312₂₃₅, TaLOX-B1a/Xwmc312₂₄₇, TaLOX-B1b/Xwmc312₂₂₇, TaLOX-B1b/Xwmc312₂₃₅ and TaLOX-B1b/Xwmc312₂₄₇, were found, with 7.2%, 9.5%, 12.1%, 23.2%, 9.5% and 38.6%, respectively, in the bread wheat cultivars surveyed.Analysis of association of LOX alleles and LOX activity indicated that the average of LOX activity of wheat cultivars with Xwmc312₂₃₅ is significantly higher than those of cultivars with Xwmc312₂₂₇ and Xwmc312₂₄₇, and the average of LOX activity of cultivars with TaLOX-B1a is significantly higher than those of cultivars with TaLOX-B1b (P＜0.05). Moreover, analysis of association of LOX allele combination and LOX activity showed that the LOX activity (76.803 AU·min^-1·g^-1) of the cultivars with TaLOX-B1a/Xwmc312₂₃₅ is significantly higher than those of other five LOX allele combinations, whereas the LOX activity (62.45 AU·min^-1·g^-1) of the cultivars with TaLOX-B1b/Xwmc312₂₂₇ genotype is significantly lower than those of other five combinations. 【Conclusion】It was concluded that LOX activities of most of the wheat cultivars from the Yellow and Huai valleys were middle type and the wheat cultivars with extremely low (less than 40 AU·min^-1·g^-1 ) or high (more than 90 AU·min^-1·g^-1 ) LOX activities were very scarce in the Yellow and Huai wheat regions. Amongst 6 different combinations of LOX genes, wheat cultivars with TaLOX-B1a/Xwmc312₂₃₅possessed relatively the highest LOX activity and wheat cultivars with TaLOX-B1b/Xwmc312₂₂₇ possessed relatively the lowest LOX activity in the Yellow and Huai wheat regions (P＜0.05). These results offered important gene germplasms for improvement of LOX activities of bread wheat in wheat breeding program. This study could also provide important information for wheat quality improvement in China.

Key words: bread wheat, flour color, lipoxygenase, allelic variation

吴培培，宋双，张福彦，陈锋，崔党群. 黄淮麦区部分小麦种质脂肪氧化酶活性分析及等位基因检测[J]. 中国农业科学, 2015, 48(2): 207-214.

WU Pei-pei, SONG Shuang, ZHANG Fu-yan, CHEN Feng, CUI Dang-qun. The Allelic Variation of Lipoxygenase Genes in Bread Wheat Cultivars from the Yellow and Huai Wheat Areas of China[J]. Scientia Agricultura Sinica, 2015, 48(2): 207-214.

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