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作物学报 ›› 2019, Vol. 45 ›› Issue (4): 546-555.doi: 10.3724/SP.J.1006.2019.84096

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

以分子标记辅助连续回交快速提高花生品种油酸含量及对其后代农艺性状的评价

黄冰艳,齐飞艳,孙子淇,苗利娟,房元瑾,郑峥,石磊,张忠信,刘华,董文召,汤丰收,张新友()   

  1. 河南省农业科学院经济作物研究所 / 农业部黄淮海油料作物重点实验室 / 河南省油料作物遗传改良重点实验室, 河南郑州 450002
  • 收稿日期:2018-07-10 接受日期:2018-12-24 出版日期:2019-04-12 网络出版日期:2019-01-04
  • 通讯作者: 张新友
  • 作者简介:huangbingyan@aliyun.com
  • 基金资助:
    本研究由国家现代农业产业技术体系建设专项(CARS-13);河南省花生产业技术体系和玛氏-中国高油酸花生育种计划项目(Mars-China HOAP2013-2018)

Improvement of oleic acid content in peanut (Arachis hypogaea L.) by marker assisted successive backcross and agronomic evaluation of derived lines

HUANG Bing-Yan,QI Fei-Yan,SUN Zi-Qi,MIAO Li-Juan,FANG Yuan-Jin,ZHENG Zheng,SHI Lei,ZHANG Zhong-Xin,LIU Hua,DONG Wen-Zhao,TANG Feng-Shou,ZHANG Xin-You()   

  1. Industrial Crops Research Institute, Henan Academy of Agricultural Sciences / Huanghuaihai Key Laboratory of Oil Crops, Ministry of Agriculture / Henan Provincial Key Laboratory for Oil Crops Improvement, Zhengzhou 450002, Henan, China
  • Received:2018-07-10 Accepted:2018-12-24 Published:2019-04-12 Published online:2019-01-04
  • Contact: Xin-You ZHANG
  • Supported by:
    This study was supported by the China Agricultural Research System(CARS-13);Henan Agricultural Research System(Mars-China HOAP2013-2018)

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摘要：

高油酸是花生重要的品质性状, 高油酸花生及其制品具有较好的品质稳定性和较高的营养和保健价值。我国高油酸花生的育成品种类型较少, 遗传背景不够丰富, 育种手段比较单一。针对上述问题, 本研究开发了AS-PCR-MP高油酸分子标记检测方法, 优化了KASP分子标记检测体系, 利用分子标记辅助连续回交, 结合近红外品质快速检测技术及南繁加代技术, 以河南省大面积推广的豫花15、远杂9102、豫花9327、豫花9326四个不同类型品种为轮回亲本, 5年内连续回交4代、自交4代, 定向获得了4个轮回亲本遗传背景的BC4F4和BC4F5稳定高油酸改良材料24个。调查分析了BC4F4和BC4F5单株的13个农艺性状与轮回亲本的相似度, 并利用轮回亲本与非轮回亲本之间的差异SNP的KASP分子标记进行了BC4F4和BC4F5株系的轮回亲本遗传背景检测。结果表明, 轮回亲本的遗传背景在BC4F5的比例为79.49%~92.31%。本研究为快速高效改良花生油酸含量探索了新的方法, 获得的新品系拓展了高油酸花生的遗传背景, 获得的一系列近等基因系可作为遗传研究材料进一步加以利用。

关键词: 花生, 高油酸, 脂肪酸脱氢酶(FAD2), 分子标记辅助回交, 遗传背景

Abstract:

High oleic acid content is a key quality trait in peanut varieties. Peanut and its products with high oleic acid content possess better quality stability and nutrition for benefiting peanut processing and human health. To date, the released high oleic acid varieties were few and most of which were derived by conventional breeding with narrow genetic background. In this study, Allele Specific-PCR-Mispairing (AS-PCR-MP) method and optimized KASP assay system were developed to facilitate successive marker-assisted backcross breeding strategy for high oleic acid content. We also applied Near Infra-Red technology and winter nursery in the five-year successive backcross and selfing. Four types of cultivars (Yuhua 15, Yuanza 9102, Yuhua 9327, and Yuhua 9326) were selected as recurrent parents for backcrossing. Twenty-four BC4F4 and BC4F5 stable lines with different genetic backgrounds and high oleic acid content were produced from four generations of successive backcrosses and four generations of BC4 selfing. Similarities between BC4 selfing progenies and recurrent parents were analyzed based on 13 agronomic traits. The recovery rate of genetic background for BC4F4 and BC4F5 was investigated by designing KASP assays with SNPs that were polymorphic between recurrent parents and non-recurrent parents. The proportion of genetic background of recurrent parents in BC4F5 was 79.49% to 92.31%. This study provides a new strategy for efficiently improving oleic acid content of peanuts with diverse genetic backgrounds. The near isogenic lines obtained in this study could be valuable genetic resources for further utilizations.

Key words: peanut (Arachis hypogaea L.), high oleic acid content, fatty acid desaturase (FAD2), marker assisted backcross (MABC), genetic background

图1

分子标记辅助连续回交高油酸花生育种路线图 RP: 轮回亲本; HO: 高油酸亲本; AS-PCR-MP: 位点特异PCR; NIR: 近红外检测。"

图2

AS-PCR-MP扩增的F1的4种基因型带型"

图3

KASP检测BC4F2单株FAD2的突变位点SNP"

表1

远杂9102 BC4F2:3后代油酸含量分布及其F2基因型"

F2:3油酸含量
F2:3 oleic acid content		 F2基因型
Related F2 genotype		 F2:3单株数量
Number of F2:3 plants		 F2基因型比例
Genotype ratio
<38% _ _ _ _*; _ol1 _ _ 86 9.4/16
38%-48% _ _ _ol2; ol1ol1 _ _ 29 3.2/16
48%-68% ol1ol1 _ol2; _ol1ol2ol2; _ol1 _ol2; __ ol2ol2 25 2.7/16
>68% ol1ol1ol2ol2 7 0.8/16
合计Total 147

图4

KASP检测BC4F4:5 FAD2的突变位点SNP"

图5

不同轮回亲本的BC4F4的性状分布箱线图 蓝色粗实线代表中位数, 黑色虚线代表轮回亲本平均数, 粉色虚线代表非轮回亲本平均数。YH15: 豫花15; YZ9102: 远杂9102; YH9327: 豫花9327; YH9326: 豫花9326。"

表2

KASP检测BC4F5株行遗传背景"

遗传背景
Genetic background		 YH15 BC4F5 lines YH15 BC4F4 lines YZ9102 BC4F5 lines YZ9102 BC4F4 lines
Z172000 Z172003 Z172025 Z172250 Z172290 Z172081 Z172086 Z172291 Z172299
轮回亲本RP (%) 79.49 84.62 92.31 66.67 77.78 87.50 87.50 83.33 83.33
杂合率Heterozygous (%) 7.69 7.69 7.69 22.22 0.00 6.25 6.25 16.67 16.67
非轮回亲本NRP (%) 12.82 7.69 0.00 11.11 22.22 6.25 6.25 0.00 0.00

表3

优良单株与亲本荚果及籽仁性状比较"

基因型
Genotype		 优良单株数(个)
No. of superior plants		 油酸含量
Oleic acid (%)		 油亚比
O/L		 荚果类型
Pod type		 荚果网纹
Pod reticulation		 种皮颜色
Testa color
豫花15
Yuhua 15		 38.71 0.96 普通型
Virginia		 粗浅
Thick and slight		 粉红色
Tan
远杂9102
Yuanza 9102		 34.50 0.81 珍珠豆型
Spanish		 细深
Thin and prominent		 粉红色
Tan
豫花9327
Yuhua 9327		 39.25 0.98 普通型
Virginia		 粗浅
Thick and slight		 粉红色
Tan
豫花9326
Yuhua 9326		 36.01 0.82 普通型
Virginia		 粗深
Thick and prominent		 粉红色
Tank
开农176
Kainong 176		 70.93 5.00 普通型
Virginia		 粗浅
Thick and slight		 粉红色
Tan
DF12 75.09 7.52 普通型
Virginia		 细浅
Thin and slight		 粉红色
Tan
开选016
Kaixuan 016		 70.03 4.96 普通型
Virginia		 粗浅
Thick and slight		 粉红色
Tan
高油酸豫花15
HOAP YH 15*		 8 70.26-74.21 5.14-10.72 普通型
Virginia		 粗浅
Thick and slight		 粉红色
Tan
高油酸远杂9102
HOAP YZ 9102*		 8 73.37-79.64 8.81-42.82 珍珠豆型
Spanish		 细深
Thin and prominent		 粉红色
Tan
高油酸豫花9327
HOAP YH 9327*		 2 71.90-76.37 6.90-11.77 普通型
Virginia		 粗浅
Thick and slight		 粉红色
Tan
高油酸豫花9326
HOAP YH 9326*		 6 73.63-82.51 6.79-34.96 普通型
Virginia		 粗深
Thick and prominent		 粉红色
Tan
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