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作物学报 ›› 2018, Vol. 44 ›› Issue (02): 208-217.doi: 10.3724/SP.J.1006.2018.00208

• • 上一篇    下一篇

我国部分主推小麦品种组织培养再生能力评价

张伟1,2,**, 尹米琦1,**, 赵佩1, 王轲1, 杜丽璞1, 叶兴国1,*   

  1. 1 中国农业科学院作物科学研究所 / 农作物基因资源与遗传改良国家重大科学工程, 北京 100081
    2 广州甘蔗糖业研究所海南甘蔗育种场, 海南三亚 572025
  • 收稿日期:2017-03-26 接受日期:2017-09-10 出版日期:2018-02-12 网络出版日期:2017-09-28
  • 通讯作者: 张伟,尹米琦,叶兴国
  • 作者简介:

    第一作者联系方式: 张莉, E-mail: windylili@foxmail.com; 荐红举, E-mail: jianhongju1989@126.com; ** 同等贡献(Contributed equally to this work)

  • 基金资助:
    本研究由国家重点研发计划项目(2016YFD0102001)和国家转基因生物新品种培育重大专项(2016ZX08010004)资助

Regeneration Capacity Evaluation of Some Largely Popularized Wheat Varieties in China

Wei ZHANG**, Mi-Qi YIN**, Pei ZHAO, Ke WANG, Li-Pu DU, Xing-Guo YE*   

  1. 1 Institute of Crop Sciences, Chinese Academy of Agricultural Sciences / National Key Facility for Gene Resources and Genetic Improvement, Beijing 100081
    2 Hainan Sugarcane Breeding Station, Guangzhou Sugarcane Industry Research Institute, Sanya 572025, Hainan, China
  • Received:2017-03-26 Accepted:2017-09-10 Published:2018-02-12 Published online:2017-09-28
  • Contact: Wei ZHANG,Mi-Qi YIN,Xing-Guo YE
  • Supported by:
    This study was supported by the National Key Research and Development Program of China (2016YFD0102001) and the National Major Project for Developing New GM Crops (2016ZX08010004).

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摘要：

小麦细胞工程育种和基因工程育种存在强烈的基因型特异性, 从目前推广的优良小麦品种中筛选不同外植体再生能力强的基因型, 对于提高小麦生物技术育种效率和加速育成品种的生产应用具有重要意义。本研究以全国大面积推广的24个优良小麦品种和抗白粉病优良品系CB037为材料, 连续2年进行花药培养、幼胚培养和成熟胚培养, 统计愈伤组织诱导率、愈伤组织分化率和植株再生率, 分析、评价这些小麦品种(系) 3种外植体的组织培养再生性能。结果表明, 25个小麦品种(系)花药、幼胚、成熟胚的植株再生率分别为0~41.75%、2.25%~531.92%和3.24%~84.34%, 基因型差异显著; 组织培养再生能力以幼胚最强(119.79%), 成熟胚其次(36.23%), 花药最弱(4.91%)。CB037的3种外植体组织培养再生效率均最高, 轮选987、扬麦16、内麦836、科农199、新春6号、郑麦366、郑麦9023、新冬20、烟农19和川麦42幼胚培养植株再生能力表现较强, 新春6号、京冬8号、石麦4185、科农199和轮选987成熟胚培养植株再生率较高, 石麦4185和邯6172花药培养绿苗诱导率较高。小麦组织培养效率与基因型和外植体类型密切相关, 不同品种同一外植体再生能力差异显著, 同一品种不同外植体再生能力也存在显著差异, 并且3种外植体的组织培养再生能力不存在相关性。本研究筛选到不同外植体再生能力较好的优良小麦基因型, 可进一步用于小麦转基因育种和单倍体育种。

关键词: 小麦, 花药, 幼胚, 成熟胚, 组织培养

Abstract:

Strong genotype dependence exists in wheat doubling haploid and genetic engineering breeding, in which high regeneration ability is a main restrain. In this study, we evaluated the regeneration abilities of various explants of 24 commercial popularized wheat varieties and a new breeding line (CB037) with high powdery mildew resistance. The explants used in the two-year experiment were anther, immature embryo, and mature embryo. The regeneration potential was assessed based on callus induction rate, callus differentiation rate, and shoot induction rate. The plantlet regeneration rates of the all genotypes tested were 0-41.75% for anther culture, 2.25%-531.92% for immature embryo culture, and 3.24%-84.34% for mature embryo culture, showing significant differences among genotypes. Generally, immature embryos had stronger regeneration ability (119.79%) than mature embryos (36.23%) and anthers (4.91%). Among all the used genotypes, CB037 showed the highest regeneration rates for the three explant types. Lunxuan 987, Yangmai 16, Neimai 836, Kenong 199, Xinchun 6, Zhengmai 366, Zhengmai 9023, Xindong 20, Yannong 19, and Chuanmai 42 exhibited ideal regeneration abilities in immature embryo culture; Xinchun 6, Jingdong 8, Shimai 4185, Kenong 199, and Luanxun 987 performed good in mature embryo culture; and Shimai 4185 and Han 6172 were characterized with high regeneration rate of green plantlet in anther culture. Plant regeneration efficiency of wheat was closely associated with genotype and explant type. The same explant type from different genotypes showed significantly different regeneration abilities, and different explants of the same genotype showed different regeneration abilities. However, there was no correlation among regeneration abilities of the three types of explants. The selected genotypes with high regeneration potential are recommended for application in genetic and cell engineering breeding of wheat.

Key words: wheat, anther, immature embryo, mature embryo, tissue culture

附表1

本研究用培养基配方"

培养基
Medium		 组成成分
Component
W14 W14 + 2 mg L-1 2,4-D + 0.5 mg L-1 KT + 100 g L-1 蔗糖 Sucrose + 2.4 g L-1 植物凝胶 Phytagel, pH 5.8
SD2 MS (不含MS维生素) + 30 g L-1 蔗糖 Sucrose + 150 mg L-1 Asp + 10.0 mg L-1 Vb1 + 2.0 mg L-1 2,4-D + 2.4 g L-1植物凝胶Phytagel, pH 5.8
Adi

 MS (不含MS维生素) + 0.75 g L-1 MgCl2 + 15 g L-1 甘露醇 Mannitol + 15 g L-1 山梨醇Sorbitol + 8 g L-1琼脂 Agar +5.0 mg L-1 谷氨酰胺 Glutamine + 0.5 g L-1 CH + 12.0 g L-1葡萄糖 Glucose + 10 mg L-1 Vb1 + 1.0 mg L-1 Vb6 + 1.0 mg L-1 烟酸 Nicotinic acid + 2.0 mg L-1 Gly + 39 mg L-1 AS + 2 mg L-1 Dicamba + 4 mg L-1 AgNO3 + 40 mg L-1 Cys + 100 mg L-1 Vc, pH 5.8
1/2MSNK 1/2MS +0.5 mg L-1 NAA+1 mg L-1 KT+20 g L-1蔗糖 Sucrose +2.4 g L-1植物凝胶 Phytagel, pH 5.8
IESDI2 MS + 30 g L-1蔗糖 Sucrose + 2.4 g L-1 植物凝胶 Phytagel + 2 mg L-1 Dicamba, pH 5.8
FHCK MS +20 g L-1蔗糖 Sucrose + 2.4 g L-1植物凝胶 Phytagel, pH 5.8

表1

不同小麦基因型幼胚培养愈伤组织诱导率、分化率和再生率比较"

图1

不同小麦基因型花药培养的愈伤组织诱导率、愈伤组织分化率、绿苗再生率和白化苗再生率"

图2

部分小麦基因型花药愈伤组织诱导和分化情况 A: 周麦18花药培养愈伤组织诱导情况; B: CB037愈伤组织分化情况; C: 周麦18愈伤分化情况; D: 石麦4185分化绿苗和白苗情况。"

图3

小麦品种CB037 (A)、扬麦16 (B)、郑麦9023 (C)和石麦4185 (D)幼胚培养植株再生情况"

图4

小麦品种CB037 (A)、轮选987 (B)、克丰10 (C)和克丰12 (D)的成熟胚培养植株再生情况"

表2

不同小麦基因型成熟胚培养愈伤组织分化率和再生率比较"

基因型
Genotype		 成熟胚数
Mature embryos		 分化愈伤组织数 Differentiated calli 再生绿苗数
Shoots regenerated		 愈伤组织分化率
Differentiated calli rate (%)		 绿苗诱导率
Shoot produced rate (%)
CB037 764 412 621 54.73 84.34
新春6号 Xinchun 6 730 426 500 58.48 68.78
京冬8号 Jingdong 8 722 402 441 55.83 61.95
石麦4185 Shimai 4185 368 188 215 51.31 59.00
科农199 Kenong 199 346 198 181 57.16 52.34
轮选 987 Lunxuan 987 368 150 190 41.05 52.19
周麦18 Zhoumai 18 584 256 254 44.17 44.36
新冬20 Xindong 20 362 136 154 37.56 42.56
扬麦16 Yangmai 16 356 138 149 39.04 42.25
郑麦366 Zhengmai 366 360 148 150 41.11 41.65
鄂麦18 Emai 18 328 138 126 43.38 40.28
周麦22 Zhoumai 22 364 146 132 40.02 36.17
中麦895 Zhongmai 895 320 108 111 34.86 35.85
川麦42 Chuanmai 42 352 112 116 31.73 32.97
济麦22 Jimai 22 366 124 117 33.73 31.82
烟农19 Yannong 19 368 154 115 42.05 31.27
宁春4号 Ningchun 4 378 106 98 28.52 26.44
西农979 Xinong 979 464 130 114 27.63 23.91
矮抗58 Aikang 58 346 90 73 25.50 20.59
邯6172 Han 6172 468 90 103 16.86 19.79
龙麦30 Longmai 30 334 80 58 23.98 17.80
郑麦9023 Zhengmai 9023 484 90 71 18.62 14.73
内麦836 Neimai 836 216 18 28 7.66 14.30
克丰12 Kefeng 12 322 20 27 6.32 7.24
克丰10号 Kefeng 10 408 20 13 5.03 3.24
LSD0.01 15.25 28.47

表3

小麦不同外植体组织培养特性间的相关系数"

外植体类型
Explant type		 愈伤组织诱导 Callus induction 愈伤组织分化 Callus differentiation 植株再生 Plant regeneration
幼胚
Immature embryo		 成熟胚
Mature embryo		 幼胚
Immature embryo		 成熟胚
Mature embryo		 幼胚
Immature embryo		 成熟胚
Mature embryo
成熟胚 Mature embryo 0.18 1.00 0.30 1.00 0.51 1.00
花药 Anther -0.13 -0.14 0.42 0.33 0.32 0.53
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