水稻半育突变体sfp10的生理特征分析及基因定位

doi:10.3724/SP.J.1006.2023.22005

作物学报 ›› 2023, Vol. 49 ›› Issue (3): 634-646.doi: 10.3724/SP.J.1006.2023.22005

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

水稻半育突变体sfp10的生理特征分析及基因定位

李秋平¹(), 张春龙¹, 杨宏¹, 王拓¹, 李娟¹, 金寿林¹, 黄大军¹, 李丹丹¹^,²^,^*(), 文建成¹^,^*()

¹云南农业大学稻作研究所, 云南昆明 650201
²云南省作物生产与智慧农业重点实验室, 云南昆明 650201

收稿日期:2022-01-21 接受日期:2022-06-07 出版日期:2023-03-12 网络出版日期:2022-07-05
通讯作者: 李丹丹,文建成
作者简介:E-mail: qiupingyouxiang@163.com
基金资助:
云南省重大科技专项(202102AE090017);云南省基础研究计划面上项目(202201AT070263);云南省教育厅科学研究基金项目(2022J0280);云南省作物生产与智慧农业重点实验室开放基金课题(20210101)

Physiological characteristics analysis and gene mapping of a semi-sterility plant mutant sfp10 in rice (Oryza sativa L.)

LI Qiu-Ping¹(), ZHANG Chun-Long¹, YANG Hong¹, WANG Tuo¹, LI Juan¹, JIN Shou-Lin¹, HUANG Da-Jun¹, LI Dan-Dan¹^,²^,^*(), WEN Jian-Cheng¹^,^*()

¹Rice Research Institute of Yunnan Agricultural University, Kunming 650201, Yunnan, China
²Key Laboratory of Crop Production and Intelligent Agriculture of Yunnan Province, Kunming 650201, Yunnan, China

Received:2022-01-21 Accepted:2022-06-07 Published:2023-03-12 Published online:2022-07-05
Contact: LI Dan-Dan,WEN Jian-Cheng
Supported by:
Major Science and Technology Project of Yunnan Province(202102AE090017);Basic Research General Program of Yunnan Province(202201AT070263);Science Research Fund Project of Yunnan Education Department(2022J0280);Open Fund Project of Key Laboratory of Crop Production and Intelligent Agriculture of Yunnan Province(20210101)

摘要/Abstract

摘要：

以水稻花粉半育性突变体lsm与籼稻93-11构建的高世代回交导入系semi-fertility plant 10 (sfp10)为研究对象, 与野生型93-11相比, 突变体在在株高、叶长、叶宽、分蘖数、花粉数量等农艺性状上均未发现显著差异, 但花粉育性却显著下降。花粉镜检及花粉发育后期的扫描电镜观察结果显示, sfp10突变体部分花粉在发育后期淀粉积累减少并最终败育。花粉发育相关生理指标检测结果表明, 突变体花药中脯氨酸和淀粉的含量显著下降; 蔗糖在突变体穗部上游组织(源叶、库叶、茎)中积累量显著增加, 但穗部含量却明显减少, 说明蔗糖到穗部的运输过程受到影响。遗传分析表明, sfp10突变体性状受1对隐性核基因控制, 基因初定位将突变位点定位于水稻10号染色体RM25389和RM25404之间的398 kb区间内, 该区间包含3个与蔗糖转运相关的基因和1个与淀粉合成相关的基因。本研究为进一步开展花粉半育性调控基因的精细定位、基因功能及调控机制的深入研究奠定基础。

关键词: 水稻, 半育性植株, sfp10, 基因定位, 淀粉积累

Abstract:

In this study, the advanced backcross population of semi-fertility plant 10 (sfp10) mutation constructed by rice pollen semi-fertile mutant lsm and indica rice cultivar 93-11 (wild-type, WT) was used as the research subjects. Compared to the WT, there were no significant differences in plant height, leaf length, leaf width, tiller number, pollen number and other agronomic traits, but pollen fertility reduced significantly. Pollen microscopy and scanning electron microscopy (SEM) at the late stage of pollen development showed that starch accumulation in some pollen of sfp10 decreased observably, eventually led to pollen abortion. Physiological indexes related pollen development revealed that the contents of proline and starch in pollen of the sfp10 mutant decreased significantly. Sucrose accumulation generally increased in the upstream tissues (source leaf, sink leaf, and stem) of panicle, but sucrose content decreased significantly in panicle, indicating that transport from sucrose to panicle was affected. Genetic analysis indicated that the sfp10 phenotype was controlled by a pair of recessive nuclear genes. Gene preliminary mapping located the mutant site into a 398 kb interval between RM25389 and RM25404 on rice chromosome 10, which contained 3 genes related to sucrose transport and 1 gene related to starch synthesis. This study laid a foundation for further research on fine mapping, gene function, and regulation mechanism of pollen semi-fertility regulation genes.

Key words: rice, semi-fertility plant, sfp10, gene mapping, starch accumulation

李秋平, 张春龙, 杨宏, 王拓, 李娟, 金寿林, 黄大军, 李丹丹, 文建成. 水稻半育突变体sfp10的生理特征分析及基因定位[J]. 作物学报, 2023, 49(3): 634-646.

LI Qiu-Ping, ZHANG Chun-Long, YANG Hong, WANG Tuo, LI Juan, JIN Shou-Lin, HUANG Da-Jun, LI Dan-Dan, WEN Jian-Cheng. Physiological characteristics analysis and gene mapping of a semi-sterility plant mutant sfp10 in rice (Oryza sativa L.)[J]. Acta Agronomica Sinica, 2023, 49(3): 634-646.

图/表 10

附表1

用于sfp10基因定位的分子标记"

标记 Marker	染色体 Chr.	正向序列 Forward sequence (5'-3')		反向序列 Reverse sequence (5'-3')	扩增长度 Amplification length (bp)
RM24899	10	CACAAGTTCTGGGTCTCAGTGG		AGAGGATCAACATCGCAATACC	280
RM24985	10	CACCTCCTCTCTCCTCTCTATCTTCC		TATAAGGGCTTTCGGATGGATGG	262
RM25003	10	GATTGATCCGAGAGACAAATCC		TCGATCAATAGTAGCAGCAGTAGG	116
RM25164	10	AAGAAGCGGACAGAAAGAAAGG		AGAAGGAACGCACCCTAACG	368
RM25741	10	AAGGCCAGGCGGATTAGTGG		AGAGGCTGGGTCCTTCCTCTCG	115
RM25810	10	GAAGGGCAGGAGGAGATAGTAGTAGG		CGTCATTGTCGCCAGAAAGC	134
RM25273	10	ATCCAAGAACCATGTCGTTTCG		TGCACAGTGGTAGCCAACTTATCC	345
RM5806	10	GAATGCTAATTGCGGTTGAAGC		GGATCTTTCCTCCCAATCTTTGC	176
RM25425	10	CCAGCCCAAACAGCTCTTGC		GGGCACTGTTTGTCTTTCTGTGC	155
mfp15	10	AGGGTAGAGTATGTCGGTGTTTCC		CCGTGGCAGTAGCAGTAGGC	180
mfp41	10	GACACGGTTCTCCTTCCACTACC		CGATGATAGCGTTAGCCATAACA	260
RM6142	10	CCTGCTTCTCCCTCCTGTACCC		GCGAGCAAATACAGAGGCTACTACC	100
RM25348	10	CTCACGCCGATCAGAGCAAGC		GCATCTCCATTCTACCACCCATTCG	283
RM25368	10	TATAGTTAAGGGAGCCACGCAAGC		CCACCTCGTAAGAACATGGAGAACC	952
RM25379	10	CCCTTAGATTCCTGCAGCTTTCC		AAGAGGGAATTGGAGGAATGAGC	216
RM25385	10	GCATCTCAAAGGAACTGACTGACC		ATGAGATGCCTGCCATTGTACC	117
RM25389	10	GGAATGGGAGCTTCTGGCTAGG		GGCGTGCACTGGCATAATTACC	471
RM25404	10	GCAACGGTTCTCCTTCCACTACC		CCATGATAGCGTTAGCCATAAACG	265
RM5708	10	TGGTATCCATAACCTTGACAGC		GTCACGTATCGAACAGTCTCC	249
RM24875	10	GATGAGCGAGGACAAGAATTAAGC		GCTAGCTGATTCAGATCCCATCG	198
RM24956	10	TTCGTGTGCATTCCCACACATCC		AAGGGTGAGGTGGCGGCTTAGG	298
RM25142	10	TGTGAGGAGAGCCATTTAGC		GGGTTCCAGTCTTGAATTTAGC	206
RM25334	10	CCGTACACCGAGCTCATCACC		CTTGGACAGCTTGAACGAGATCC	212
RM25359	10	CCTGCCGAAACCAACCTTATTCC		GCGAAGGTAAAGCCCAATACTGTTCC	212
RM25362	10	CAGGAGCTCGCGGTTCATCG		AGAGGAGGTTCTCCCTCGTGTGC	77
RM25365	10	TCTCGTCAACTTCGCTTGTATCG		CGATTCGACTCTAAACAGTGATGC	155
RM25366	10	TCGGTCTCTGTGCCGTGATTAGG		CACCAGCGCAGCAACTAACATCC	105
RM25375	10	TGTAGCTGCACATCTCCTTCAGC		GCTCATCTCCAAGCTGCAGTCC	218
RM25376	10	GCGCTGACCTTCGGCACTTAGC		CGTTCCGGATTTATCTGCTTGAGACC	447
RM25377	10	TTGGTTTCCTAGCTTGGCCTACG		TAGAATGCGATGCGAGACAGTGG	160
RM25378	10	CCGTACGTTCAGGTATGTGTTTAAGG		AGCAAACAGGCAGCATAAGAGG	400
RM25380	10	CGAGAACAGCTCCGAGATCAACC		ATGGAGGCGTAGAACGGGATGC	111
RM8201	10	CCCACTATGCTGGTACACATCTTTCG		CCTCTTCCGCTTCGCCTTCG	291
RM25386	10	GAGCACGAAAGCATGGGACAACC		CGCAGAGCTCCAAGAAACACAGC	214
RM6868	10	GAGGTGAACATGCCGAGGAAGC		GGCCGGAGTATATAGAACCCAAAGC	188
标记 Marker	染色体 Chr.		正向序列 Forward sequence (5'-3')	反向序列 Reverse sequence (5'-3')	扩增长度 Amplification length (bp)
RM25407	10		TAGAAAGCCCACTTCCAACACG	CCAGCTCGATACCCAAGTAAGTCC	400
RM14	1		CCGAGGAGAGGAGTTCGAC	GTGCCAATTTCCTCGAAAAA	191
RM486	1		CCCCCCTCTCTCTCTCTCTC	CCCCCCTCTCTCTCTCTCTC	104
RM315	1		GAGGTACTTCCTCCGTTTCAC	AGTCAGCTCACTGTGCAGTG	133
RM579	1		TCCGAGTGGTTATGCAAATG	AATTGTGTCCAATGGGCTGT	182
RM23	1		CATTGGAGTGGAGGCTGG	GTCAGGCTTCTGCCATTCTC	145
RM211	2		CCGATCTCATCAACCAACTG	CTTCACGAGGATCTCAAAGG	161
RM112	2		GGGAGGAGAGGCAAGCGGAGAG	AGCCGGTGCAGTGGACGGTGAC	128
RM110	2		TCGAAGCCATCCACCAACGAAG	TCCGTACGCCGACGAGGTCGAG	156
RM475	2		CCTCACGATTTTCCTCCAAC	ACGGTGGGATTAGACTGTGC	235
RM6	3		GTCCCCTCCACCCAATTC	TCGTCTACTGTTGGCTGCAC	163
RM156	3		GCCGCACCCTCACTCCCTCCTC	TCTTGCCGGAGCGCTTGAGGTG	160
RM514	3		AGATTGATCTCCCATTCCCC	CACGAGCATATTACTAGTGG	259
RM293	3		TCGTTGGGAGGTATGGTACC	CTTTATCTGATCCTTGGGAAGG	207
RM143	3		GTCCCGAACCCTAGCCCGAGGG	AGAGGCCCTCCACATGGCGACC	207
RM523	3		AAGGCATTGCAGCTAGAAGC	GCACTTGGGAGGTTTGCTAG	148
RM503	3		CACCTTTCACACACACACAC	GCCCCACTAACAAAACCAAG	268
RM401	4		TGGAACAGATAGGGTGTAAGGG	CCGTTCACAACACTATACAAGC	283
RM471	4		ACGCACAAGCAGATGATGAG	GGGAGAAGACGAATGTTTGC	106
RM273	4		GAAGCCGTCGTGAAGTTACC	GTTTCCTACCTGATCGCGAC	207
RM334	5		GTTCAGTGTTCAGTGCCACC	GACTTTGATCTTTGGTGGACG	182
RM163	5		ATCCATGTGCGCCTTTATGAGGA	CGCTACCTCCTTCACTTACTAGT	124
RM440	5		CATGCAACAACGTCACCTTC	ATGGTTGGTAGGCACCAAAG	169
RM136	6		GAGAGCTCAGCTGCTGCCTCTAGC	GAGGAGCGCCACGGTGTACGCC	101
RM204	6		GTGACTGACTTGGTCATAGGG	GCTAGCCATGCTCTCGTACC	169
RM589	6		ATCATGGTCGGTGGCTTAAC	CAGGTTCCAACCAGACACTG	186
RM412	6		CACTTGAGAAAGTTAGTGCAGC	CCCAAACACACCCAAATAC	198
RM527	6		GGCTCGATCTAGAAAATCCG	TTGCACAGGTTGCGATAGAG	233
RM427	7		TCACTAGCTCTGCCCTGACC	TGATGAGAGTTGGTTGCGAG	185
RM432	7		TTCTGTCTCACGCTGGATTG	AGCTGCGTACGTGATGAATG	187
RM38	8		ACGAGCTCTCGATCAGCCTA	TCGGTCTCCATGTCCCAC	250
RM296	9		CACATGGCACCAACCTCC	GCCAAGTCATTCACTACTCTGG	123
RM206	11		CCCATGCGTTTAACTATTCT	CGTTCCATCGATCCGTATGG	147
RM224	11		ATCGATCGATCTTCACGAGG	TGCTATAAAAGGCATTCGGG	157
RM254	11		AGCCCCGAATAAATCCACCT	CTGGAGGAGCATTTGGTAGC	165
RM473	11		TATCCTCGTCTCCATCGCTC	AAGGATGTGGCGGTAGAATG	97
RM28778	12		CTTCATCACCGCCTCCGTTCC	AGCTCTCTCCCGCTCTGGATGC	165
RM28802	12		GGAGGCTTAACTCAGCACTACTGG	CATGCTCAGATGTGTTCACTTGG	179
RM28812	12		GGGAATTGAGAATCGACAGAAACC	CCAGTACGTCAAACAGGGCTACG	195
RM28819	12		GAACGTCTCGTTCCCTATCACG	TCCACTCACTCATCTCTCCTTGC	244
RM27545	12		GCAGGTTAGTTCACTCCATGTGC	TTCGAAGGGTCTTGTGATGTATCC	375

附表1

图1

表1

图2

附表2

图3

表2

图4

图5

表3

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性状 Trait	野生型93-11 Wild type 93-11	突变体sfp10 sfp10 mutant	相比93-11 Compared with 93-11 (%)
株高Plant height (cm)	109.10±4.75	111.10±4.23	1.83
分蘖数Tiller number	5.00±1.81	4.35±0.93	-13.00
剑叶长Swordleaf length (cm)	37.60±1.36	38.63±2.01	2.49
剑叶宽Swordleaf width (cm)	1.39±0.06	1.38±0.07	-0.72
生物产量Biological yield (g)	29.88±7.71	21.27±7.27	-28.81^**
经济产量Economic yield (g)	12.37±3.26	7.98±2.74	-35.47^**
结实率Seed setting rate (%)	92.84±2.65	59.34±8.79	-36.09^**
千粒重1000-grain weight (g)	29.63±2.09	25.60±2.43	-13.58^**
主穗粒数Grain number per panicle	131.15±20.08	60.40±11.86	-53.95^**

时期 Stage	花粉长度Pollen grain length (µm)		花粉宽度Pollen grain width (µm)
时期 Stage	93-11	sfp10	93-11	sfp10
六期 Stage 6	32.9	30.8	31.2	29.3
	32.2	32.7	29.8	29.0
	32.0	31.2	29.2	28.5
P值 P-value	0.28		0.15
七期 Stage 7	39.9	40.7	38.8	37.4
	40.4	39.9	39.1	38.9
	40.2	37.7	39.8	38.7
P值 P-value	0.46		0.18
八期 Stage 8	39.3	44.3	38.7	43.7
	44.1	43.1	43.2	42.6
	44.6	43.6	45.3	42.2
P值 P-value	0.59		0.84

组合 Combination	F₂		χ²_(3:1)	χ²_0.05
组合 Combination	野生型株数 No. of wild type plants	突变型株数 No. of mutant type plants	χ²_(3:1)	χ²_0.05
Sfp10/93-11	297	210	72.03	3.84
93-11/Sfp10	439	47	60.09	3.84
合计Total	736	257	0.37

参与过程 Participation process	基因名称 Gene name	基因注释 Gene annotation
氨基酸合成与代谢 Amino acid synthesis and metabolism	Os10g0405100	Similar to serine/threonine-protein kinase NAK
	Os10g0400100	Methionyl-tRNA synthetase, class Ia domain containing protein
	Os10g0406300	Ascorbate oxidase promoter-binding protein
	Os10g0400800	Similar to phenylalanyl-tRNA synthetase alpha chain
蔗糖转运 Sucrose transport	Os10g0404500	Similar to sucrose transporter
	Os10g0404533	Similar to sucrose transport protein SUT3
	Os10g0404566	Similar to sucrose transporter
淀粉合成Starch synthesis	Os10g0390500	Starch synthesis in developing seeds

水稻半育突变体sfp10的生理特征分析及基因定位

Physiological characteristics analysis and gene mapping of a semi-sterility plant mutant sfp10 in rice (Oryza sativa L.)

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