中国水稻科学 ›› 2023, Vol. 37 ›› Issue (1): 45-54.DOI: 10.16819/j.1001-7216.2023.220109
唐杰1(), 龙湍2, 吴春瑜1, 李新鹏1, 曾翔1, 吴永忠1, 黄培劲1
收稿日期:
2022-01-09
修回日期:
2022-03-22
出版日期:
2023-01-10
发布日期:
2023-01-10
通讯作者:
唐杰
基金资助:
TANG Jie1(), LONG Tuan2, WU Chunyu1, LI Xinpeng1, ZENG Xiang1, WU Yongzhong1, HUANG Peijin1
Received:
2022-01-09
Revised:
2022-03-22
Online:
2023-01-10
Published:
2023-01-10
Contact:
TANG Jie
摘要: 目的 雄性不育是水稻杂种优势利用的基础。为进一步解析其调控机制,对一个雄性不育突变体进行了鉴定。方法 利用60Co-γ对籼稻品种93-11进行辐射诱变,从其后代中鉴定到一个雄性不育突变体tms3650。将籼稻明恢63作父本同突变体tms3650杂交构建F2和F3群体,采用图位克隆的方法对目的基因进行精细定位。结果 tms3650突变体其他农艺性状与野生型一致,但花药白绿且瘦小,花粉不能被1%碘-碘化钾溶液染成蓝黑色,穗子包颈,抽穗期延迟。遗传分析表明该突变体表型受一对隐性核基因控制。精细定位结果表明基因位于第3染色体长臂SSR标记RM15927和RM15934之间135.25 kb距离内,且与RM15931标记共分离。陵水冬季南繁鉴定发现,该突变体育性受光温环境影响,说明tms3650突变体的雄性育性在短日低温条件下发生了转育,是一个光温敏雄性不育突变体。结论 通过将定位位点与已报道的雄性不育基因比较,发现tms3650是一个新的基因位点,暂命名为TMS3650。
唐杰, 龙湍, 吴春瑜, 李新鹏, 曾翔, 吴永忠, 黄培劲. 水稻光温敏雄性不育突变体tms3650的鉴定和基因定位[J]. 中国水稻科学, 2023, 37(1): 45-54.
TANG Jie, LONG Tuan, WU Chunyu, LI Xinpeng, ZENG Xiang, WU Yongzhong, HUANG Peijin. Identification and Gene Mapping of a New Photo-thermo-sensitive Male Sterile Mutant tms3650 in Rice[J]. Chinese Journal OF Rice Science, 2023, 37(1): 45-54.
图1 野生型93-11和突变体tms3650表型鉴定 A?野生型93-11(左)和突变体tms3650(右)整株;B?野生型93-11(左)和突变体tms3650(右)的穗;C?野生型93-11(左)和突变体tms3650(右)小穗表型;D?野生型93-11(左)和突变体tms3650(右)花药表型;E?野生型93-11成熟花粉碘染结果;F?突变体tms3650成熟花粉碘染结果;G?突变体tms3650成熟花粉陵水转育后碘染结果。
Fig. 1. Phenotypic comparison of wild type 93-11 (WT)and tms3650. A, Wild type 93-11 (left) and tms3650 (right); B, Panicles of wild type 93-11 (left) and mutant tms3650 (right); C, Spikelets of wild type 93-11 (left) and mutant tms3650 (right); D, Anther of wild type 93-11(left) and mutant tms3650 (right); E, Pollen fertility of wild type; F, Pollen fertility of tms3650; G, Pollen fertility of mutant tms3650 after transplanting in Lingshui, Hainan.
育性 Fertility | 株系Line | 合计 Total | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
L1 | L2 | L3 | L4 | L5 | L6 | L7 | L8 | L9 | ||
可育花粉数Fertile pollen | 170 | 174 | 156 | 120 | 202 | 71 | 95 | 100 | 39 | 1127 |
不育花粉数Sterile pollen | 195 | 215 | 271 | 130 | 190 | 85 | 104 | 81 | 16 | 1287 |
自交收种Selfed seed | 240 | 200 | 300 | 320 | 380 | 117 | 194 | 220 | 90 | 2061 |
结实率Seed setting rate/% | 32.0 | 26.6 | 40.0 | 42.6 | 50.6 | 15.6 | 25.8 | 29.3 | 12.0 | 30.5 |
表1 突变体tms3650株系育性转换分析
Table 1. Fertility conversion analysis of mutant tms3650 lines.
育性 Fertility | 株系Line | 合计 Total | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
L1 | L2 | L3 | L4 | L5 | L6 | L7 | L8 | L9 | ||
可育花粉数Fertile pollen | 170 | 174 | 156 | 120 | 202 | 71 | 95 | 100 | 39 | 1127 |
不育花粉数Sterile pollen | 195 | 215 | 271 | 130 | 190 | 85 | 104 | 81 | 16 | 1287 |
自交收种Selfed seed | 240 | 200 | 300 | 320 | 380 | 117 | 194 | 220 | 90 | 2061 |
结实率Seed setting rate/% | 32.0 | 26.6 | 40.0 | 42.6 | 50.6 | 15.6 | 25.8 | 29.3 | 12.0 | 30.5 |
图2 野生型93-11和突变体tms3650品种真实性鉴定 泳道1~4为野生型93-11;泳道5~8为突变体tms3650。
Fig. 2. Authenticity identification of wild-type 93-11 and mutant tms3650. Lanes 1-4 are wild-type 93-11; Lanes 5-8 are mutant tms3650.
杂交组合 Cross combination | F1结实率 Seed-setting rate of F1/% | F2 | χ2 3:1 | χ20.05 | |
---|---|---|---|---|---|
野生型 Wild type plant | 突变体 Mutant plant | ||||
tms3650/93-11 | 58.13 | 79 | 31 | 0.44 | 3.841 |
tms3650/ZH11 | 76.92 | 56 | 14 | 0.70 | 3.841 |
tms3650/明恢63 tms3650/Minghui 63 | 75.19 | 186 | 61 | 0.01 | 3.841 |
表2 突变体tms3650的遗传分析
Table 2. Genetic analysis of mutant tms3650.
杂交组合 Cross combination | F1结实率 Seed-setting rate of F1/% | F2 | χ2 3:1 | χ20.05 | |
---|---|---|---|---|---|
野生型 Wild type plant | 突变体 Mutant plant | ||||
tms3650/93-11 | 58.13 | 79 | 31 | 0.44 | 3.841 |
tms3650/ZH11 | 76.92 | 56 | 14 | 0.70 | 3.841 |
tms3650/明恢63 tms3650/Minghui 63 | 75.19 | 186 | 61 | 0.01 | 3.841 |
图3 tms3650突变体的精细定位 A?突变基因初步定位在第3染色体;B?突变基因精细定位在135.25 kb区间;C?定位区间内24个候选基因。
Fig. 3. Fine mapping of tms3650 mutants. A, Preliminary mapping of TMS3650 on chromosome 3; B, Fine mapping of TMS3650 in the 135.25 kb range; C, Predicted 24 candidate genes.
序号 Serial number | 基因 Gene | 功能注释 Function annotation |
---|---|---|
1 | LOC_Os03g53570 | 反转录转座子蛋白 Retrotransposon protein, putative, unclassified, expressed |
2 | LOC_Os03g53580 | 表达蛋白 Expressed protein |
3 | LOC_Os03g53590 | 表达蛋白 Expressed protein |
4 | LOC_Os03g53600 | HTH DNA结合蛋白 HTH DNA-binding protein, putative, expressed |
5 | LOC_Os03g53610 | 晚期胚胎发育丰富的蛋白质 D-34 late embryogenesis abundant protein D-34, putative, expressed |
6 | LOC_Os03g53620 | 晚期胚胎发育丰富的蛋白质 D-34 late embryogenesis abundant protein D-34, putative, expressed |
7 | LOC_Os03g53630 | PHD锌指家族蛋白 PHD finger family protein, putative, expressed |
8 | LOC_Os03g53640 | 表达蛋白 Expressed protein |
9 | LOC_Os03g53650 | 半胱氨酸合酶 Cysteine synthase, putative, expressed |
10 | LOC_Os03g53660 | 含有肌球蛋白头部结构域的蛋白质 Myosin head domain containing protein, expressed |
11 | LOC_Os03g53670 | 含有YT521-B样家族结构域的蛋白质 YT521-B-like family domain containing protein, expressed |
12 | LOC_Os03g53680 | 表达蛋白 Expressed protein |
13 | LOC_Os03g53690 | 氧化还原酶,含有短链脱氢酶/还原酶家族结构域的蛋白质 Oxidoreductase, short chain dehydrogenase/reductase family domain containing protein, expressed |
14 | LOC_Os03g53700 | PHD锌指家族蛋白 PHD-finger domain containing protein, putative, expressed |
15 | LOC_Os03g53710 | 醛糖1-差向异构酶 Aldose 1-epimerase, putative, expressed |
16 | LOC_Os03g53720 | SRPK4蛋白 SRPK4, putative, expressed |
17 | LOC_Os03g53730 | 黄素蛋白wrbA Flavoprotein wrbA, putative, expressed |
18 | LOC_Os03g53740 | 表达蛋白 Expressed protein |
19 | LOC_Os03g53750 | 核前蛋白A识别因子 Nuclear prelamin A recognition factor, putative, expressed |
20 | LOC_Os03g53760 | 核前蛋白A识别因子 Nuclear prelamin A recognition factor, putative, expressed |
21 | LOC_Os03g53770 | 含有蛋白质的RNA识别基序 RNA recognition motif containing protein, putative, expressed |
22 | LOC_Os03g53780 | 铵转运体2 Ammonium transporter 2, putative, expressed |
23 | LOC_Os03g53790 | 周质β-葡萄糖苷酶前体 Periplasmic beta-glucosidase precursor, putative, expressed |
24 | LOC_Os03g53800 | 周质β-葡萄糖苷酶前体 Periplasmic beta-glucosidase precursor, putative, expressed |
表3 目标区域预测基因
Table 3. Predicted gene in target region.
序号 Serial number | 基因 Gene | 功能注释 Function annotation |
---|---|---|
1 | LOC_Os03g53570 | 反转录转座子蛋白 Retrotransposon protein, putative, unclassified, expressed |
2 | LOC_Os03g53580 | 表达蛋白 Expressed protein |
3 | LOC_Os03g53590 | 表达蛋白 Expressed protein |
4 | LOC_Os03g53600 | HTH DNA结合蛋白 HTH DNA-binding protein, putative, expressed |
5 | LOC_Os03g53610 | 晚期胚胎发育丰富的蛋白质 D-34 late embryogenesis abundant protein D-34, putative, expressed |
6 | LOC_Os03g53620 | 晚期胚胎发育丰富的蛋白质 D-34 late embryogenesis abundant protein D-34, putative, expressed |
7 | LOC_Os03g53630 | PHD锌指家族蛋白 PHD finger family protein, putative, expressed |
8 | LOC_Os03g53640 | 表达蛋白 Expressed protein |
9 | LOC_Os03g53650 | 半胱氨酸合酶 Cysteine synthase, putative, expressed |
10 | LOC_Os03g53660 | 含有肌球蛋白头部结构域的蛋白质 Myosin head domain containing protein, expressed |
11 | LOC_Os03g53670 | 含有YT521-B样家族结构域的蛋白质 YT521-B-like family domain containing protein, expressed |
12 | LOC_Os03g53680 | 表达蛋白 Expressed protein |
13 | LOC_Os03g53690 | 氧化还原酶,含有短链脱氢酶/还原酶家族结构域的蛋白质 Oxidoreductase, short chain dehydrogenase/reductase family domain containing protein, expressed |
14 | LOC_Os03g53700 | PHD锌指家族蛋白 PHD-finger domain containing protein, putative, expressed |
15 | LOC_Os03g53710 | 醛糖1-差向异构酶 Aldose 1-epimerase, putative, expressed |
16 | LOC_Os03g53720 | SRPK4蛋白 SRPK4, putative, expressed |
17 | LOC_Os03g53730 | 黄素蛋白wrbA Flavoprotein wrbA, putative, expressed |
18 | LOC_Os03g53740 | 表达蛋白 Expressed protein |
19 | LOC_Os03g53750 | 核前蛋白A识别因子 Nuclear prelamin A recognition factor, putative, expressed |
20 | LOC_Os03g53760 | 核前蛋白A识别因子 Nuclear prelamin A recognition factor, putative, expressed |
21 | LOC_Os03g53770 | 含有蛋白质的RNA识别基序 RNA recognition motif containing protein, putative, expressed |
22 | LOC_Os03g53780 | 铵转运体2 Ammonium transporter 2, putative, expressed |
23 | LOC_Os03g53790 | 周质β-葡萄糖苷酶前体 Periplasmic beta-glucosidase precursor, putative, expressed |
24 | LOC_Os03g53800 | 周质β-葡萄糖苷酶前体 Periplasmic beta-glucosidase precursor, putative, expressed |
图5 连锁标记RM15931亲本多态性鉴定 泳道1~9分别为tms3650、野香B、93-11、明恢63、GD-7S、隆科638S、博II B、H28B、R51084。
Fig. 5. Identification of parental polymorphism of linked markers RM15931. Lanes 1-9 are tms3650, Yexiang B, 93-11, Minghui 63, GD-7S, Longke 638S, Bo II B, H28B, R51084.
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