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作物学报 ›› 2017, Vol. 43 ›› Issue (06): 855-861.doi: 10.3724/SP.J.1006.2017.00855

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

利用回交法快速选育高油酸花生新品系

于明洋1,孙明明1,郭悦1,姜平平1,雷永2,黄冰艳3,冯素萍4,郭宝珠5,隋炯明1,王晶珊1,乔利仙1,*   

  1. 1 青岛农业大学生命科学学院 / 山东省高校植物生物技术重点实验室, 山东青岛 266109；2 中国农科院油料作物研究所，湖北武汉 430062；3 河南省农业科学院经济作物研究所，河南郑州 450002；4 海南热带海洋学院生命科学与生态学院，海南三亚 572022；5 USDA-Agricultural Research Service, Tifton, GA, 31793-074, USA
  • 收稿日期:2016-04-06 修回日期:2017-01-21 出版日期:2017-06-12 网络出版日期:2017-02-17
  • 通讯作者: 乔利仙，E-mail: lxqiao73@163.com, Tel: 0532-86080640
  • 基金资助:

    本研究由国家自然科学基金项目(31471524, 31571705), Mars-中国高油酸育种计划项目和山东省科技发展计划项目(2014GNC110002)资助。

Breeding New Peanut Line with High Oleic Acid Content Using Backcross Method

YU Ming-Yang1,SUN Ming-Ming1,GUO Yue1,JIANG Ping-Ping1,LEI Yong2,HUANG Bing-Yan3,FENG Su-Ping4,GUO Bao-Zhu5,SUI Jiong-Ming1,WANG Jing-Shan1,QIAO Li-Xian1,*   

  1. 1 Key Lab of Plant Biotechnology in Universities of Shandong Province / College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, China; 2 Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan 430062, China; 3 Industrial Crops Research Institute, Henan Academy of Agricultural Sciences, Zhengzhou 450002, China; 4 College of Life Sciences and ecology, Hainan Tropical Ocean University, Sanya 572022, China; 5 USDA-Agricultural Research Service, Tifton, GA, 31793-074, USA
  • Received:2016-04-06 Revised:2017-01-21 Published:2017-06-12 Published online:2017-02-17
  • Contact: Qiao Lixian，E-mail: lxqiao73@163.com, Tel: 0532-86080640
  • Supported by:

    This program was financially supported by the National Natural Science Foundation of China (31471524, 31571705), the Mars-China High Oleic Acid Peanut Breeding Project, and the Shandong Province Science and Technology Development Plan Project (2014GNC110002).

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摘要：

以普通花生品种花育22为母本、高油酸花生品种开农176为父本杂交得到F1杂种，筛选油酸含量高于60%且同时含有FAD2a和FAD2b位点的F1为杂交父本，以花育22为轮回亲本(母本）连续回交得到BC1F1~BC4F1代回交种。利用近红外光谱仪测定F1及BC1F1~BC4F1籽粒的油酸、亚油酸含量，选择油酸含量大于60%的种子，用刀片切取种子小部分子叶提取DNA，以F0.7/R3为引物进行PCR扩增及测序，根据测序峰图差异表现筛选出同时含有FAD2a和FAD2b位点的种子作为下一代回交的父本。切去部分子叶的种子切口用石蜡封闭，播种前浸泡于40℃温水中催芽，对12 h后未露白的种子用100 mg L–1乙烯利浸泡4 h后再转入40℃温水浸泡至24 h，发芽率可达到98%。2013年春季开始杂交，2016年春在青岛播种BC4F2代种子，取幼苗期幼叶鉴定基因型，筛选出基因型为aabb的单株，收获时选留农艺性状类似于花育22的优良单株，再利用近红外光谱仪测定所选单株油酸含量，获得油酸含量在70%以上、油酸亚油酸比值大于7.0的单株24个。这些单株与花育22相比，农艺性状基本相同，称为改良花育22高油酸花生新品系。

关键词: 花生, 高油酸, 回交, 测序法, F0.7/R3

Abstract:

F1 hybrids and BC1F1–BC4F1 backcross generation were obtained by cross and backcross with peanut variety HY22 as female parent (recurrent parent) and Kainong176 with high oleic acid content as the donor parent. The contents of oleic acid and linoleic acid of F1 and BC1F1 to BC4F1 were determined by Near infrared spectrometer. The seeds with oleic acid content higher than 60% were selected and their partial cotyledon was cut off for DNA extraction. The loci FAD2a and FAD2b in these seeds were detected by sequencing absorption peaks of PCR products with F0.7/R3 as primers, and those seeds with both FAD2a and FAD2b were used as the male parent in following backcross. The incision in these seeds whose cotyledon was cut off was sealed by paraffin oil, and these seeds were soaked in water for sprouting at 40°C. Those seeds that did not germinate within 12 hours were soaked in 100 mg L–1 ethephon solution for four hours, then immersed into water till 24 hours at 40°C by which 98% seeds could germinate. Four to five times of backcross every two years were finished in spring in Qingdao, 1–2 times in autumn and winter in Sanya, by which the breeding process might be accelerated. The backcrossing work began in the spring of 2013 and BC4F2 seeds were sowed in Qingdao in the spring of 2016. The young leaves were used for genotyping and these plants with aabb genotype and similar agronomic characters with Huayu 22 were selected and harvested, whose oleic content was then confirmed by NIR. Twenty single plants with similar characters of Huayu 22 whose oleic content was higher than 70% and oleic/linoleic ratio was higher than 7.0 were regarded as a new improved HY22 strain with high oleic acid content.

Key words: Peanut (Arachis hypogaea L.), High oleic acid content, Backcross, Sequencing, F0.7/R3

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