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作物学报 ›› 2023, Vol. 49 ›› Issue (8): 2039-2050.doi: 10.3724/SP.J.1006.2023.22057

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

四倍体水稻回复二倍体品系的籼粳属性鉴定和杂种优势利用初探

宋兆建1,*(), 冯紫旖1, 屈天歌1, 吕品苍1, 杨晓璐1, 湛明月1, 张献华1, 何玉池1, 刘育华2, 蔡得田1,2,*()   

  1. 1 湖北大学生命科学学院, 湖北武汉430062
    2 武汉多倍体生物科技有限公司, 湖北武汉430345
  • 收稿日期:2022-09-27 接受日期:2023-02-10 出版日期:2023-08-12 网络出版日期:2023-02-28
  • 通讯作者: 宋兆建,蔡得田
  • 作者简介:E-mail: zjsong99@126.com
  • 基金资助:
    湖北省重点研发计划项目(2020BBA032);武汉市重大科技专项和武汉市品牌农业发展计划项目资助

Indica-japonica attribute identification and heterosis utilization of diploid rice lines reverted from tetraploid rice

SONG Zhao-Jian1,*(), FENG Zi-Yi1, QU Tian-Ge1, LYU Pin-Cang1, YANG Xiao-Lu1, ZHAN Ming-Yue1, ZHANG Xian-Hua1, HE Yu-Chi1, LIU Yu-Hua2, CAI De-Tian1,2,*()   

  1. 1 School of Life Sciences, Hubei University, Wuhan 430062, Hubei, China
    2 Wuhan Polyploid Biotechnology Co., Ltd., Wuhan 430345, Hubei, China
  • Received:2022-09-27 Accepted:2023-02-10 Published:2023-08-12 Published online:2023-02-28
  • Contact: SONG Zhao-Jian,CAI De-Tian
  • Supported by:
    Key Research and Development Program of Hubei Province(2020BBA032);Major Science and Technology Project of Wuhan, and the Brand Agriculture Development Plan of Wuhan

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摘要：

本研究以8个四倍体水稻回复二倍体品系为材料, 采用InDel分子标记法鉴定其籼粳属性; 同时以回复二倍体品系为父本, 分别与籼稻光温敏雄性不育系培矮64S、粳稻光温敏雄性不育系农垦58S配制杂交组合, 进行亲本及杂种的遗传距离及聚类分析, 考察杂种及其父本的产量相关性状, 分析杂种的超父本优势、超标优势; 另外, 对亲本遗传距离与杂种优势的相关性进行分析, 探讨利用亲本遗传距离预测杂种优势的可行性。结果表明: (1) 回复二倍体品系的籼型基因频率分布在0.605~0.947之间, 所有品系都兼有籼稻和粳稻遗传成分, 但籼粳成分比例各不相同。(2) 回复二倍体品系和培矮64S之间的遗传距离在0.21~0.42之间, 回复二倍体品系和农垦58S之间的遗传距离在0.68~0.95之间; 籼粳属性相同或相近的材料聚为一类, 清晰显示了材料间的亲缘关系。(3) 回复二倍体品系的杂种优势利用需要广亲和基因的参与, 以具有广亲和基因的培矮64S为母本配制的杂交组合在单株有效穗数、每穗总粒数和每穗实粒数方面超父本优势和超标优势明显; 在综合后的单株产量上8个杂交组合都表现出超父本优势和超标优势, 尤其是杂交组合HYP2单株产量超标优势突出, 高达45.92%。(4) 亲本遗传距离与杂种F1的每穗总粒数和单株产量呈显著正相关, 说明InDel遗传距离可以用于杂种优势预测。研究证实了籼粳亚种间四倍体水稻的回复二倍体品系兼有籼稻和粳稻遗传成分, 遗传多样性丰富, 利用这些回复二倍体品系能够配制出超强杂种优势组合, 为通过以籼粳中间型品系与广亲和光温敏雄性不育系配组实现籼粳亚种间杂种优势利用提供了理论依据和参考; 同时初步证实了“以多倍体为变异载体选育回复二倍体水稻”育种途径的可行性, 为水稻育种提供了新的思路和途径。

关键词: 水稻, 多倍体, 回复二倍体, InDel, 籼粳属性, 遗传距离, 杂种优势

Abstract:

In this study, the indica-japonica attribute of eight reverted diploid lines were identified by InDel molecular marker. The hybrid combinations were made between the reverted diploid lines and photoperiod- and thermo-sensitive genic male sterile lines Peiai 64S (indica) and Nongken 58S (japonica), respectively. The genetic distance and cluster analysis of the parents and hybrids were carried out, the yield related traits of the hybrids and the male parents were investigated, then the over male parent heterosis and the competitive heterosis of the hybrids were analyzed. In addition, the correlation between parental genetic distance and the heterosis was analyzed to explore the feasibility of using parental genetic distance to predict heterosis. The results showed as follows: (1) The indica gene frequency of the reverted diploid lines ranged from 0.605 to 0.947. All lines had both indica and japonica genetic components, but the proportions of indica and japonica components were different from each other. (2) The genetic distance between the reverted diploid lines and Pei’ai 64S were 0.21-0.42, and that between the reverted diploid lines and Nongken 58S were 0.68-0.95. The materials with the same or similar indica-japonica attribute were clustered together, which clearly showed the genetic relationship between the materials. (3) The heterosis utilization of the reverted diploid lines needed the participation of wide compatibility genes. The hybrid combinations made between Peiai 64S that had wide compatibility gene and the reverted diploid lines had obvious over male parent heterosis and competitive heterosis in the effective panicle number per plant, the total grain number per panicle, and the filled grain number per panicle. In the yield per plant, eight hybrid combinations all showed over male parent heterosis and competitive heterosis, especially the yield per plant of HYP2 was prominent, up to 45.92%. (4) The genetic distance of parents was significantly positively correlated with the total grain number per panicle and the yield per plant of the hybrids, indicating that the InDel genetic distance can be used for predicting heterosis. The study confirmed that the reverted diploid lines from the tetraploid indica-japonica subspecies rice had both indica and japonica genetic components, and contained abundant genetic diversity. Using the reverted diploid lines, the hybrid combinations with super heterosis could be made, which providing a theoretical basis and reference for the utilization of indica-japonica subspecies heterosis through the combination of indica-japonica intermediate lines and photoperiod- and thermo-sensitive genic male sterile lines with wide compatibility genes. At the same time, we preliminarily confirmed the feasibility of the pathway “breeding new diploid rice varieties by using polyploid as the vector for creating variation”, which provided a new idea and approach for rice breeding.

Key words: rice, polyploid, reverted diploid, InDel, indica-japonica attribute, genetic distance, heterosis

表1

用于籼粳属性鉴定的19对InDel分子标记及序列"

标记名称
Marker		 正向引物DNA序列
Forward sequence (5′-3′)		 反向引物DNA序列
Reverse sequence (5′-3′)		 9311与日本晴片段差异
Fragment differences between 9311 and Nipponbare (bp)
R1M7 ATTCCTGGTTCTACATTACTTA CGCCTCACTAGAATATCGGA 37
R1M37 ATAGTTCGCCATCGTCAT ACACGCCATAGCAAGGAA 53
R2M10 CCCAGTCTGCTGCCATCT GAATGTATTTCAGTTCCAGTAAG 48
R2M50 CCTGAAGGAAATGATAGCAATAG GTTTTGTATGCTCTTCACTTGTC 42
R3M10 CCGAGTACCATTGCTTTC CTGCCATAGTTACTGCTCTGTT 37
R3M53 ACACTGGCTACGGCAAAG TTTGTTCGGGAATAATGATGC 35
R4M17 AGTGCTCGGTTTTGTTTTC GTCAGATATAATTGATGGATGTA 51
R5M13 GAGAAAGAGTGGAAGGAG AGTATCGTCAGGAGGGTC 32
R5M30 CTCAATTTCACCCATCCC CGCTCCGTCTCCAACCTC 46
R6M14 AAATGTCCATGTGTTTGCTTC CATGTGTGGAATGTGGTTG 34
R6M44 TTAGGAATAAAGGCTGGATA TTACCGTTAATAGGTGGAA 34
R7M7 ACCTTCCCTCCCCTTTTGAT AACTTGGTCTTCCTGTTTTATTG 67
R7M37 CAGCCCTAAATCTAAATACCC ACGTTGAGACAGGCGAGC 36
R8M33 CGAAAGAGGAGAGGGGTAGT CGAAAACGAGAAACAAATA 38
R9M42 CTATAAGACCAAAACGAAAACT GAAAACCATTGTGTCACTGTA 48
R10M17 TGAACAATAAACCACAGAAGCA CCCTTTATTCCCTCCTTTG 31
R11M23 AAGGTTGACAAGGACAGAAG TCGCAGGAATGGATAAAA 42
R12M10 ATCATTTCAGCCTGTGCC AGCTTAATAGGGGGGACG 47
R12M27 ATTTCATTGCCATCAGTT GTAATCTTCTATCCGTTCA 33

表2

依据籼型(Fi)或粳型(Fj)基因频率鉴定籼粳属性的InDel分类标准"

籼粳属性
Indica-japonica
attributes		 基因频率 Gene frequency
籼型(Fi) Indica 粳型(Fj) Japonica
典型籼稻Typical indica 0.90-1.00 0-0.10
籼稻Indica 0.75-0.89 0.11-0.25
偏籼Indica cline 0.61-0.74 0.26-0.39
中间类型Intermediate 0.40-0.60 0.40-0.60
偏粳Japonica cline 0.26-0.39 0.61-0.74
粳稻Japonica 0.11-0.25 0.75-0.89
典型粳稻Typical japonica 0-0.10 0.90-1.00

表3

回复二倍体品系、不育系及其杂种的籼粳属性鉴定结果"

材料名称
Material		 材料类型
Type		 籼型基因频率(Fi)
Gene frequencies of indica		 粳型基因频率(Fj)
Gene frequencies of japonica		 籼粳属性鉴定结果
Indica-japonica attributes
identified by InDel markers
9311 籼稻对照品种Control variety of indica 1.000 0.000 典型籼稻Typical indica
日本晴Nipponbare 粳稻对照品种Control variety of japonica 0 1.000 典型粳稻Typical japonica
A3 回复二倍体Reverted diploid line 0.658 0.342 偏籼Indica cline
T1 回复二倍体Reverted diploid line 0.605 0.395 偏籼Indica cline
ST110 回复二倍体Reverted diploid line 0.868 0.132 籼稻Indica
ST112 回复二倍体Reverted diploid line 0.947 0.053 典型籼稻Typical indica
ST126 回复二倍体Reverted diploid line 0.789 0.211 籼稻Indica
ST130 回复二倍体Reverted diploid line 0.816 0.184 籼稻Indica
ST133 回复二倍体Reverted diploid line 0.789 0.211 籼稻Indica
ST141 回复二倍体Reverted diploid line 0.895 0.105 籼稻Indica
PA64S 籼稻不育系Indica sterile line 0.842 0.158 籼稻Indica
NK58S 粳稻不育系Japonica sterile line 0.105 0.895 粳稻Japonica
HYP1 杂交组合Hybrid rice 0.737 0.263 偏籼Indica cline
HYP2 杂交组合Hybrid rice 0.737 0.263 偏籼Indica cline
HYP3 杂交组合Hybrid rice 0.816 0.184 籼稻Indica
HYP4 杂交组合Hybrid rice 0.816 0.184 籼稻Indica
HYP5 杂交组合Hybrid rice 0.737 0.263 偏籼Indica cline
HYP6 杂交组合Hybrid rice 0.763 0.237 籼稻Indica
HYP7 杂交组合Hybrid rice 0.711 0.289 偏籼Indica cline
HYP8 杂交组合Hybrid rice 0.816 0.184 籼稻Indica
HYN1 杂交组合Hybrid rice 0.447 0.553 中间类型Intermediate
HYN2 杂交组合Hybrid rice 0.447 0.553 中间类型Intermediate
HYN3 杂交组合Hybrid rice 0.289 0.711 偏粳Japonica cline
HYN4 杂交组合Hybrid rice 0.289 0.711 偏粳Japonica cline
HYN5 杂交组合Hybrid rice 0.447 0.553 中间类型Intermediate
HYN6 杂交组合Hybrid rice 0.316 0.684 偏粳Japonica cline
HYN7 杂交组合Hybrid rice 0.316 0.684 偏粳Japonica cline
HYN8 杂交组合Hybrid rice 0.289 0.711 偏粳Japonica cline

表4

回复二倍体品系与不育系间的遗传距离"

父本
(回复二倍体品系)
Male parent		 与母本(不育系)的遗传距离
Genetic distance between male and female parents
PA64S NK58S
A3 0.37 0.74
T1 0.42 0.68
ST110 0.26 0.95
ST112 0.21 0.89
ST126 0.32 0.89
ST130 0.21 0.89
ST133 0.26 0.89
ST141 0.26 0.95

图1

基于InDel分子标记的遗传相似系数聚类图 A: 亲本及杂种聚类图(母本培矮64S); B: 亲本及杂种聚类图(母本农垦58S)。"

表5

杂交组合及其父本的主要农艺性状"

材料/组合
Material/
combination		 生育期PD (d) 株高
PH (cm)		 单株有效穗数EP 每穗总粒数TG 每穗实粒数FG 结实率
SSR (%)		 千粒重
TGW (g)		 单株产量
YPP (g)
A3 118 135.14±1.42 11.50±1.22 308.05±11.83 267.25±12.73 86.76±2.52 26.08±0.68 80.15±11.37
T1 123 133.64±1.93 12.50±1.58 305.70±8.18 254.95±8.57 83.40±0.91 25.70±0.77 81.89±9.70
ST110 140 124.28±2.02 12.40±1.43 207.37±9.20 184.33±10.96 88.89±1.91 27.06±0.67 61.85±8.57
ST112 136 123.24±2.19 11.60±1.51 210.55±14.88 186.51±12.74 88.58±2.54 30.80±0.48 66.63±8.94
ST126 130 123.82±1.30 14.20±1.14 269.44±23.02 229.79±16.62 85.29±3.93 22.48±0.56 73.35±7.16
ST130 133 124.45±1.93 13.60±1.35 270.00±22.36 232.65±16.22 86.17±2.61 23.53±0.69 74.44±10.83
ST133 135 118.62±1.44 13.80±1.48 246.94±16.64 217.76±14.75 88.18±1.11 25.55±0.49 76.78±10.29
ST141 132 130.03±1.83 11.60±1.07 207.96±9.77 193.62±8.68 93.10±1.58 30.12±0.78 67.65±6.17
HYP1 124 140.14±1.18 14.30±1.34 338.16±22.07 277.86±12.07 82.17±3.65 23.74±0.57 94.33±7.53
HYP2 130 139.36±1.12 15.20±0.92 384.31±8.87 312.76±9.94 81.38±2.68 23.69±0.33 112.63±9.81
HYP3 137 128.64±1.47 15.40±1.07 289.04±12.75 253.96±13.55 87.86±2.39 25.51±0.50 99.76±8.21
HYP4 139 125.06±1.13 14.40±0.84 277.27±15.42 225.31±13.98 81.26±2.08 26.58±0.41 86.23±9.49
HYP5 136 127.74±0.69 14.80±1.14 370.73±20.99 301.37±15.72 81.29±0.93 22.85±0.60 101.93±9.35
HYP6 135 134.64±1.38 14.00±1.41 335.93±11.54 305.76±12.46 91.02±2.44 22.61±0.40 96.78±12.05
HYP7 140 129.83±1.87 13.40±1.35 338.17±18.98 309.46±16.91 91.51±1.04 23.75±0.61 98.48±13.52
HYP8 135 131.77±1.35 13.90±0.99 265.30±14.24 222.73±13.38 83.95±2.02 27.16±0.62 84.10±10.89
HYN1 135 97.23±2.35 29.50±1.65 92.28±5.40 13.32±2.37 14.13±1.86 26.52±1.48 10.34±1.79
HYN2 132 101.97±2.36 32.00±2.54 105.31±8.24 27.16±3.45 25.71±2.15 25.32±1.26 21.88±3.71
HYN3 142 96.35±2.29 36.60±2.01 161.26±6.80 18.58±2.09 11.80±1.26 24.37±1.48 17.13±3.14
HYN4 145 87.93±2.51 25.20±1.81 127.12±2.52 11.46±1.00 8.70±0.73 25.06±1.44 6.89±1.03
HYN5 140 80.08±2.32 33.50±2.80 108.46±5.04 7.79±0.62 7.25±0.56 26.72±1.36 7.27±0.49
HYN6 137 100.30±2.94 36.50±2.27 107.89±5.67 15.60±1.87 14.81±1.59 25.26±1.29 14.95±2.03
HYN7 140 101.00±2.76 33.30±2.45 166.40±10.39 13.15±1.50 7.55±0.61 24.62±0.75 10.56±1.43
HYN8 140 97.79±2.34 23.20±1.87 101.15±7.16 7.82±0.92 7.90±0.61 25.08±1.13 4.61±0.76
丰两优4号(对照)
Fengliangyou 4 (CK)		 135 132.19±1.39 10.60±1.17 278.94±11.35 245.76±13.68 88.10±2.39 29.63±0.56 77.19±10.45

表6

各杂交组合产量相关性状超父本优势的描述性统计"

统计数值
Statistical value		 单株有效穗数
EP		 每穗总粒数
TG		 每穗实粒数
FG		 结实率
SSR		 千粒重
TGW		 单株产量
YPP
均值Mean 14.80 29.14 25.62 -2.78 -6.91 33.43
极差Range 27.25 29.61 38.14 15.46 15.36 43.62
最大值Max. 24.35 39.39 42.11 5.63 1.66 61.30
最小值Min. -2.90 9.78 3.97 -9.83 -13.70 17.68
正向优势组合数PDC 7 8 8 2 1 8
负向优势组合数NDC 1 0 0 6 7 0

图2

杂交组合产量相关性状的超父本优势 EP: 单株有效穗数; TG: 每穗总粒数; FG: 每穗实粒数; SSR: 结实率; TGW: 千粒重; YPP: 单株产量。"

表7

杂交组合产量相关性状超标优势的描述性统计(对照丰两优4号)"

统计数值
Statistical value		 单株有效穗数
EP		 每穗总粒数
TG		 每穗实粒数
FG		 结实率
SSR		 千粒重
TGW		 单株产量
YPP
均值Mean 36.08 16.46 12.37 -3.46 -17.36 25.38
极差Range 18.86 42.66 36.63 11.64 15.38 36.97
最大值Max. 45.28 37.77 27.26 3.87 -8.32 45.92
最小值Min. 26.42 -4.89 -9.37 -7.77 -23.70 8.95
正向优势组合数PDC 8 6 6 2 0 8
负向优势组合数NDC 0 2 2 6 8 0

图3

杂交组合产量相关性状的超标优势(对照丰两优4号) EP: 单株有效穗数; TG: 每穗总粒数; FG: 每穗实粒数; SSR: 结实率; TGW: 千粒重; YPP: 单株产量。"

表8

亲本InDel遗传距离与产量相关性状杂种优势的相关系数"

性状
Trait		 亲本遗传距离
GD		 单株有效穗数
EP		 每穗总粒数
TG		 每穗实粒数
FG		 结实率
SSR		 千粒重
TGW
单株有效穗数EP 0.554
每穗总粒数TG 0.761* 0.198
每穗实粒数FG 0.536 -0.008 0.920**
结实率SSR -0.468 -0.460 -0.098 0.298
千粒重TGW -0.527 -0.005 -0.880** -0.941** -0.241
单株产量YPP 0.736* 0.529 0.847** 0.806* 0.010 -0.680
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