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Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (02): 208-217.doi: 10.3724/SP.J.1006.2018.00208

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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 Online:2018-02-12 Published: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).

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

Compositions of the media used in this study for callus induction and differentiation

Correlation coefficient between tissue culture traits of different wheat explants"

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

Table1

Callus induction, differentiation and plantlet regeneration of the immature embryo tissues derived from different wheat genotypes"

Fig. 1

Callus induction rate, differential calli rate, green plantlet regeneration rate, and albino plantlet regeneration rate from anther culture in different wheat genotypes"

Fig. 2

Callus induction, differentiation and plantlet regeneration of anther culture of several wheat genotypes A: callus induction from anther culture of Zhoumai 18; B: albino plantlet regeneration of Zhoumai 18; C: green plantlet regeneration of CB037; D: albino plantlet regeneration and green plantlet regeneration of Shimai 4185."

Fig. 3

Plant regeneration from immature embryos in wheat genotypes CB037 (A), Yangmai 16 (B), Zhengmai 9023 (C), and Shimai 4185 (D)"

Fig. 4

Plant regeneration from the mature embryos of wheat genotypes CB037 (A), Lunxuan 987 (B), Kefeng 10 (C), and Kefeng 12 (D)"

Table 2

Callus differentiation and plantlet regeneration from mature embryo tissues in different wheat genotypes"

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

Table 3

Correlation coefficient between tissue culture traits of different wheat explants"

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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