欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2009, Vol. 35 ›› Issue (5): 802-808.doi: 10.3724/SP.J.1006.2009.00802

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘蓝自交不亲和基因MLPK与SSP的FISH定位

荣小营1，朱利泉1,*，王 永1，高启国2，陈晓丹1，杨洋2，王小佳2   

  1. 1西南大学植物生理生物化学实验室，重庆400716；2西南大学重庆市蔬菜学重点实验室，重庆400716
  • 收稿日期:2008-10-15 修回日期:2009-01-20 出版日期:2009-05-12 网络出版日期:2009-03-23
  • 通讯作者: 朱利泉,023-68250794
  • 基金资助:

    本研究由国家自然科学基金项目（30671429），重庆市自然基金项目（9266）资助。

Localization of MLPK and SSP Genes for Self-Incompatibility of Brassica oleracea by Fluorescence in situ Hybridization

RONG Xiao-Ying1,ZHU Li-Quan1*,WANG Yong1,GAO Qi-Guo2,CHEN Xiao-Dan1,YANG Yang2,WANG Xiao-Jia2   

  1. 1Plant Physiology and Biochemistry Laboratory of Southwest University,Chongqing 400716,China;2Key Laboratory in Olericulture of Chongqing,Southwest University,Chongqing 400716,China
  • Received:2008-10-15 Revised:2009-01-20 Published:2009-05-12 Published online:2009-03-23
  • Contact: ZHU Li-Quan,023-68250794

PDF

2061

被引次数

13

摘要：

利用FISH技术, 对自交不亲和基因MLPK与SSP在甘蓝有丝分裂前中期染色体、减数分裂早粗线期染色体以及伸长DNA纤维等3种分辨率水平的靶DNA载体上进行物理定位。结果表明, 在有丝分裂前中期, MLPK探针信号位于一对近中着丝粒同源染色体的短臂中部, 距着丝粒的百分距离约为53.41±3.16；SSP探针信号位于一对具有随体的近端着丝粒同源染色体的长臂端部, 距着丝粒的百分距离约为78.36±4.26。综合3种载体上的FISH结果表明, MLPK与SSP在甘蓝染色体组中可能都只有一个同源序列座位, 具有在单倍体基因组中的单拷贝性。重复FISH杂交表明, MLPK与5S rDNA位于同一对染色体。依据Armstrong的核型分析标准, 初步判断MLPK与SSP分别位于甘蓝的2号和7号染色体, 与S位点不存在连锁关系。另从比较基因组学角度对定位结果进行了讨论。

关键词: 甘蓝, MLPK基因, SSP基因, 荧光原位杂交, 自交不亲和性

Abstract:

In Brassica, self-incompatibility recognition is controlled by the multiallelic gene complex (S-haplotypes) at the S-locus, which encodes both the male determinant S-locus pollen coat protein (SCR/SP11) and the female determinant S-locus receptor kinase (SRK). Studies of MLPK and SSP genes for self-incompatibility of Brassica have been gradually widespread since they were identified. However, the position and copy number of MLPK and SSP genes in Brassica oleracea genome are still unclear. In this paper, the localization of MLPK and SSP genes for self-incompatibility of Brassica oleracea on prometaphase chromosomes, early pachytene chromosomes and extended DNA fibers was conducted successfully by fluorescence in situ hybridization. The results indicated that MLPK probe was hybridized onto the short arm of a pair of homologous prometaphase chromosomes, and the percent distance from centromere to the signal point was about 53.41±3.16; SSP probe was hybridized onto the long arm of a pair of homologous prometaphase chromosomes with the satellite, and the percent distance from centromere to the signal point was about 78.36±4.26. Hybridization signals from three kinds of cytological targets with different FISH resolutions showed that both MLPK gene and SSP gene might be located at a single-copy locus in Brassica oleracea genome. Repeated FISH indicated that both MLPK and 5S rDNA probes were hybridized onto the same chromosomes. According to karyotype standard of Armstrong, it was primarily inferred that MLPK gene was located on the chromosome 2, and SSP gene on the chromosome 7. The results presumably revealed thatneither MLPK nor SSP is linked to S-locus. And they locate respectively on the different chromosomes in Brassica oleracea. In addition, the collinearity relationship of MLPK as well as SSP between Brassica and Arabidopsis thaliana was also discussed on the basis of comparative genomics.

Key words: Brassica oleracea, MLPK gene, SSP gene, Fluorescence in situ hybridization, Self-incompatibility


[1] Stein J C, Howlett B, Boyes D C, Nasrallah M E, Nasrallah J B. Molecular cloning of a putative receptor protein kinase gene encoded at the self-incompatibility locus of Brassica oleracea. Proc Natl Acad Sci USA, 1991, 88: 8816–8820

[2] Goring D R, Rothstein S J. The S-locus receptor kinase gene in a self-incompatible Brassica napus line encodes a functional serine/threonine kinase. Plant Cell, 1992, 4: 1273–1281

[3] Takayama S, Shiba H, Iwano M, Shimosato H, Che F S, Watanabe M, Suzuki G, Hinata K, Isogai A. The pollen determinant of self-incompatibility in Brassica campestris. Proc Natl Acad Sci USA, 2000, 97: 1920–1925

[4] Boyes D C, Nasrallah M E, Vrebalov J, Nasrallah J B. The self-incompatibility (S) haplotypes of Brassica contain highly divergent and rearranged sequences of ancient origin. Plant Cell, 1997, 9: 237–247

[5] Gu T S, Mazzurco M, Sulaman W, Matias D D, Goring D R. Binding of an arm repeat protein to the kinase domain of the S-locus receptor kinase. Proc Natl Acad Sci USA, 1998, 95: 382–387

[6] Cabrillac D, Cock J M, Dumas C, Gaude T. The S-locus receptor kinase is inhibited by thioredoxins and activated by pollen coat proteins. Nature, 2001, 410: 220–223

[7] Goring D R, Walker J C. Self-rejection — A new kinase connection. Science, 2004, 303: 1474–1475

[8] Murase K, Shiba H, Iwano M, Che F S, Watanabe M, Isogai A, Takayama S. A membrane-anchored protein kinase involved in Brassica self-incompatibility signaling. Science, 2004, 303: 1516–1519

[9] Kakita M, Murase K, Iwano M, Matsumoto T, Watanabe M, Shiba H, Isogai A, Takayama S. Two distinct forms of M-locus protein kinase localize to the plasma membrane and interact directly with S-locus receptor kinase to transduce self-incompatibility signaling in Brassica rapa. Plant Cell, 2007, 19: 3961–3973

[10] Wu N-B(吴能表). Study on evocable protein phosphorylation and correlative characters in the self-incompatible of Brassica oleracea L. PhD Dissertation of Southwest Agricultural University, 2003. pp 51–52(in Chinese with English abstract)

[11] Zhao Y-B(赵永斌), ZHU L-Q(朱利泉). Cloning and sequences analysis of SRK substrate protein gene from Brassica oleracea. Acta Agric Boreali-occident Sin (西北农业学报), 2007, 16(6): 102–106(in Chinese with English abstract)

[12] Hasterok R, Wolny E, Hosiawa M, Kowalczyk M, Kulak-Ksiazczyk S, Ksiazczyk T, Heneen W K, Maluszynska J. Comparative analysis of rDNA distribution in chromosomes of various species of Brassicaceae. Ann Bot, 2006, 97: 205–216

[13] Chen R-Y(陈瑞阳), Song W-Q(宋文芹), Li X-L(李秀兰). Wall degradation hypotonic methed of preparing chromosome sample in plant and its significance in the cytogenetics, Acta Genet Sin (遗传学报), 1982, 9(2): 151–159(in Chinese with English abstract)

[14] Zhong X-B(钟筱波), Fransz P F, Wennekes J, Kammen A, De Jong J H, Zabel P. Fluorescence in situ hybridization to pachytene chromosomes and extended DNA fibres in plants. Acta Genet Sin (遗传学报), 1998, 25(2): 142–149(in Chinese with English abstract)

[15] Yang K, Qi H Y, Zhu L Q, Wang X J. Localization of S genes on extended DNA fibers (EDFs) in Brassica oleracea by high-resolution FISH. Acta Genet Sin, 2006, 33(3): 277–284

[16] Zhao Y-B赵永斌), Zhu L-Q(朱利泉), Wang X-J(王小佳). Cloning and sequences analysis of M-locus protein kinase gene from Brassica oleracea. Acta Agron Sin (作物学报), 2006, 32(1): 46–50(in Chinese with English abstract)

[17] Cheng Z K, Buell C R, Wing R A, Jiang J M. Resolution of fluorescence in situ hybridization mapping on rice mitotic prometaphase chromosomes, meiotic pachytene chromosomes and extended DNA fibers. Chrom Res, 2002, 10: 379–387

[18] Armstrong S J, Fransz P, Marshall D F, Jones G H. Physical mapping of DNA repetitive sequences to mitotic and meiotic chromosomes of Brassica oleracea var. alboglabra by fluorescence in situ hybridization. Heredity, 1998, 81: 666–673

[19] Li G, Gao M, Yang B, Quiros C F. Gene for gene alignment between the Brassica and Arabidopsis genomes by direct transcriptome mapping. Theor Appl Genet, 2003, 107: 168–180

[20] Suwabe K, Tsukazaki H, Iketani H, Hatakeyama K, Kondo M, Fujimura M, Nunome T, Fukuoka H, Hirai M, Matsumoto S. Simple sequence repeat-based comparative genomics between Brassica rapa and Arabidopsis thaliana: The genetic origin of clubroot resistance. Genetics, 2006, 173: 309–319

[21] Lukens L, Zou F, Lydiate D, Parkin I, Osborn T. Comparison of a Brassica oleracea genetic map with the genome of Arabidopsis thaliana. Genetics, 2003, 164: 359–372

[22] Howell E C, Barker G C, Jones G H, Kearsey M J, King G J, Kop E P, Ryder C D, Teakle G R, Vicente J G, Armstrong S J. Integration of the cytogenetic and genetic linkage maps of Brassica oleracea. Genetics, 2002, 161: 1225–1234

[23] Wang C J R, Harper L, Cande W Z. High-resolution single-copy gene fluorescence in situ hybridization and its use in the construction of a cytogenetic map of maize chromosome 9. Plant Cell, 2006, 18: 529–544

[24] Kamisugi Y, Nakayama S, O’Neil C M, Mathias R J, Trick M, Fukui K. Visualization of the Brassica self-incompatibility S-locus on identified oilseed rape chromosomes. Plant Mol Biol, 1998, 38: 1081–1087

[25] Van De Rijke F M, Florijn R J, Tanke H J, Raap A K. DNA fiber-FISH staining mechanism. J Histochem Cytochem, 2000, 48: 743–745

[26] Stone S L, Anderson E M, Mullen R T, Goring DR. ARC1 is an E3 ubiquitin ligase and promotes the ubiquitination of proteins during the rejection of self-incompatible Brassica pollen. Plant Cell, 2003, 15: 885–898
[1] 左同鸿, 张贺翠, 曾静, 朱利泉. 甘蓝自交不亲和相关基因BoPUB3L的克隆与表达分析[J]. 作物学报, 2026, 52(6): 1698-1710.
[2] 王楚锐, 李开祥, 赵志, 肖麓, 唐国永, 赵志刚, 徐亮, 杜德志, 柳海东. 甘蓝型春油菜早花基因BnCRY2功能位点KASP标记的开发及应用[J]. 作物学报, 2026, 52(3): 708-721.
[3] 马毅娜, 吴晓明玉, 李藕琪, 王圆, 陈丽, 张盈川, 赵伦, 文静, 傅廷栋, 沈金雄. Bna-miR1040-EIF3A模块调控油菜开花时间的功能研究[J]. 作物学报, 2026, 52(2): 349-362.
[4] 王彬, 蒙姜宇, 邱浩良, 贺亚军, 钱伟. 甘蓝型油菜BnaDUF579基因家族的鉴定与表达模式分析[J]. 作物学报, 2025, 51(8): 2100-2110.
[5] 夏琦, 郭滢, 王坤美, 王思忆, 巨建业, 彭雅雯, 刘忠松, 夏石头. 甘蓝型油菜种子和种皮中水杨酸含量与原花色素积累的关系研究[J]. 作物学报, 2025, 51(5): 1189-1197.
[6] 王晓琳, 刘忠松, 康雷, 杨柳. 甘蓝型油菜角果长度和每角粒数基因定位以及角果皮转录组动态分析[J]. 作物学报, 2025, 51(4): 888-899.
[7] 张琴, 戴成, 马朝芝. 生长素响应报告基因转化甘蓝型油菜及各组织GUS动态信号分析[J]. 作物学报, 2025, 51(3): 667-675.
[8] 孙程明, 周晓婴, 陈锋, 张维, 王晓东, 彭琦, 郭月, 高建芹, 胡茂龙, 付三雄, 张洁夫. 长链非编码RNA (lncRNA)在甘蓝型油菜分枝角度调控中的功能分析与预测[J]. 作物学报, 2025, 51(3): 559-567.
[9] 黄绒, 周渠晨, 陈楚铭, 罗倩, 易东, 杜常欢, 黄祥宇, 盛锋, 杜雪竹. 过表达BnNRT2.3-like对油菜氮素吸收利用及产量的影响[J]. 作物学报, 2025, 51(12): 3184-3197.
[10] 巨建业, 杨柳, 陈浩, 康雷, 夏石头, 刘忠松. 单细胞核转录组分析揭示油菜种皮分化过程和种子颜色差异原因[J]. 作物学报, 2025, 51(11): 2860-2874.
[11] 张雯, 李玉, 王创, 石磊, 丁广大. 甘蓝型油菜磷转运蛋白BnaPT48的功能研究[J]. 作物学报, 2025, 51(11): 2983-2995.
[12] 胡志康, 舒雨, 王会, 杨莹莹, 廖俊宇, 刘佳, 成洪涛, 郭晨, 张园园, 刘胜毅, 胡琼, 梅德圣, 李超. 甘蓝型油菜苗期耐碱性种质综合鉴定与评价[J]. 作物学报, 2025, 51(10): 2681-2692.
[13] 王晨, 贺丹, 姚敏, 邱萍, 何昕, 熊兴华, 康雷, 刘忠松, 钱论文. 基于转录组分析鉴定甘蓝型油菜开花候选基因以及BnaCOR27功能验证[J]. 作物学报, 2025, 51(10): 2693-2704.
[14] 徐林珊, 郜耿东, 王宇, 王家星, 杨吉招, 武亚瑞, 张宵寒, 常影, 李真, 谢雄泽, 龚德平, 王晶, 葛贤宏. 甘蓝型油菜漆酶基因家族成员表达模式及与茎秆抗折力的关联分析[J]. 作物学报, 2025, 51(1): 134-148.
[15] 李嘉欣, 黄莹, 吴潞梅, 赵伦, 易斌, 马朝芝, 涂金星, 沈金雄, 傅廷栋, 文静. 甘蓝型油菜BnaSLY1基因进化分析及功能研究[J]. 作物学报, 2025, 51(1): 44-57.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备15008789号-2