Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (19): 3899-3908.doi: 10.3864/j.issn.0578-1752.2012.19.001

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS •     Next Articles

Screening and Identification of Susceptibility and Fertility Related Mutants from Transgenic Rice IRBB13 (PXa13:GFP)

 HAN  Kuan-De, KONG  Ling-Guang, JU  Yan-Hu, CHEN  Ming, CAO  Zhi-Gang, DING  Xin-Hua, CHU  Zhao-Hui   

  1. 1.山东农业大学植物保护学院/作物生物学国家重点实验室/山东省农业微生物重点实验室,山东泰安 271018
    2.黄冈市农业科学院水稻研究所,湖北黄冈 438000
  • Received:2012-05-21 Online:2012-10-01 Published:2012-07-12

Abstract: 【Objective】The objective of this study is to establish a simple and efficient mode of rice mutants selection, and lay the first stone for cloning rice genes involved in regulating xa13/ Xa13 gene expression or their mediated disease resistant /susceptible and pollen development signal pathway. 【Method】Using EMS treatment, a rice mutation population was constructed with IRBB13 (PXa13:GFP) which carrying green fluorescent protein(GFP) derived with dominant Xa13 gene promoter. Then the mutants were screened out with the following three periods: At seedling stage, selecting mutants in which GFP gene expression decline either in root or bud or non-expression; At adult stage, selecting mutants restored susceptible to PXO99; At booting stage, selecting mutants reduced fertility. 【Result】In total 3 000 M2 lines, 14 and 9 mutants were identified for both root and bud fluorescent declined and fluorescent normal only in root at seedling stage. At adult stage, relying on phenotype of PXO99 inoculation, a total of 25 lines were characterized to restore the susceptibility to PXO99. At booting stage, 7 mutants were identified, of which fertility declined and the green fluorescence of the anthers also disappeared and one mutant was found to restore susceptibility.【Conclusion】A simple and efficient mode of rice mutants selection system was established. Combining the abnormal expression pattern of GFP mutants, restored susceptible phenotype mutants and reduced fertility mutants, it was preliminarily indicated that the two signal transduction pathways mediated by xa13/ Xa13 genes are not only completely independent but also partially crossing between resistance/susceptibility and pollen development. Furthermore, after analysis and cloned the genes from these mutants, they will help us better understanding the functional mechanism of xa13/ Xa13 genes.

Key words: rice, EMS mutants, GFP, pollen development, bacterial blight

[1]Mew T W. Current status and future prospects of research on bacterial blight of rice. Annual Review of Phytopathology, 1987, 25: 359-382.

[2]Nino-Liu D O, Ronald P C, Bogdanove A. Xanthomonas oryzae pathovars: Model pathogens of a model crop. Molecular Plant Pathology, 2006, 7: 303-324.

[3]章  琦. 水稻白叶枯病抗性基因鉴定进展及其利用. 中国水稻科学, 2005, 19(5): 453-459.

Zhang Q. Highlights in identification and application of resistance genes to bacterial blight. Chinese Journal of Rice Science, 2005, 19(5): 453-459. (in Chinese)

[4]Yoshimura S, Yamanouchi U, Katayose Y, Toki S, Wang Z X, Kono I, Kurata N, Yano M, Iwata N, Sasaki T. Expression of Xa1, a bacterial blight resistance gene in rice, is induced by bacterial inoculation. Proceedings of the National Academy of Sciences of the USA, 1998, 95: 1663-1668.

[5]Song W Y, Wang G L, Chen L L, Kim H S, Pi L Y, Holsen T, Gardner J, Wang B, Zhai W X, Zhu L H, Fauquet C, Ronald P. A receptor kinase-like protein encoded by rice disease resistance gene Xa21. Science, 1995, 270: 1772-1804.

[6]Gu K, Yang B, Tian D, Wu L, Wang D, Sreekala C, Yang F, Chu Z, Wang G L, White F F. R gene expression induced by a type-III effector triggers disease resistance in rice. Nature, 2005, 435: 1122-1125.

[7]Iyer A S, McCouch S R. The rice bacterial blight resistance gene xa5 encodes a novel form of disease resistance. Molecular Plant-Microbe Interactions, 2004, 17(12): 1348-1354.

[8]Chu Z H, Yuan M, Ge X J, Yuan B, Xu C G, Li X H, Fu B Y, Li Z K, Bennetzen J L, Zhang Q F, Wang S P. Promoter mutations of an essential gene for pollen development results in disease resistance in rice. Genes and Development, 2006, 20: 1250-1255.

[9]Liu Q, Yuan M, Zhou Y, Li X H, Xiao J H, Wang S P. A paralog of the MtN3/saliva family recessively confers race-specific resistance to Xanthomonas oryzae in rice. Plant, Cell and Environment, 2011, 34: 1958-1969.

[10]Yang B, Sugio A, White F F. Os8N3 is a host disease susceptibility gene for bacterial blight of rice. Proceedings of the National Academy of Sciences of the USA, 2006, 103: 10503-10508.

[11]Greene E A, Codomo C A, Taylor N E, Henikoff J G, Till B J, Reynolds S H, Enns L C, Burtner C, Johnson J E, Odden A R, Comai L, Henikoff S. Spectrum of chemically induced mutations from a large-scale reverse-genetic screen in Arabidopsis. Genetics, 2003, 164(2): 731-740.

[12]Menda N, Semel Y, Peled D, Eshed Y, Zamir D. In silico screening of a saturated mutation library of tomato. The Plant Journal, 2004, 38(5): 861-872.

[13]Ray S K, Rajeshwari R, Sonti R V. Mutants of Xanthomonas oryzae pv. oryzae deficient in general secretory pathway are virulence deficient and unable to secrete xylanase. Molecular Plant-Microbe Interactions, 2000, 13: 394-401.

[14]Zhu Y, Zhao H F, Ren G D, Yu X F, Cao S Q, Kuai B K. Characterization of a novel developmentally retarded mutant (drm1) associated with the autonomous flowering pathway in Arabidopsis. Cell Research, 2005, 15(2): 133-140.

[15]Chu Z H, Fu B Y, Yang H, Xu C G, Li Z K, Sanchez A, Park Y J, Bennetzen J L, Zhang Q F ,Wang S P. Targeting xa13, a recessive gene for bacterial blight resistance in rice. Theoretical and Applied Genetics, 2006, 112: 455-461.

[16]Patrick R, Sabine R, Tina S, Janett E, Sebastian S, Jens B, Wang S P, Thomas L. Promoter elements of rice susceptibility genes are bound and activated by specific TAL effectors from the bacterial blight pathogen, Xanthomonas oryzae pv. Oryzae. New Phytologist, 2010, 187(4): 1048-1057.

[17]White F F, Yang B. Host and pathogen factors controlling the rice-Xanthomonas oryzae interaction. Plant Physiology, 2009, 150: 1677-1686.

[18]Jens B, Heidi S, Schornack S, Angelika L, Simone H, Sabine K, Thomas L, Anja N, Ulla B. Breaking the code of DNA binding specificity of TAL-type III effectors. Science, 2009, 326: 1509-1512.

[19]Yuan M, Chu Z, Li X, Xu C, Wang S P. Pathogen-induced expressional loss of function is key factor of race-specific bacterial resistance conferred by a recessive R gene xa13 in rice. Plant Cell Physiology, 2009, 50: 947-955.        

[20]Yuan M, Chu Z, Li X, Xu C, Wang S P. The bacterial pathogen xanthomonas oryzae overcomes rice defenses by regulating host copper redistribution. The Plant Cell, 2010, 22: 3164-3176.

[21]Antony G, Zhou J H, Sheng H, Li T, Liu B, White F F, Yang B. xa13 recessive resistance to bacterial blight is defeated by the induction of disease susceptibility gene Os11N3. The Plant Cell, 2010, 22(11): 3864-3876.

[22]Chen L Q, Hou B H, Lalonde S, Takanaga H, Hartung M L, Qu X Q, Guo W J, Kim J G, Underwood W, Chaudhuri B, Chermak D, Antaony G, White F F, Somerville S, Mudgett M B, Frommer W B. Sugar transporters for intercellular exchange and nutrition of pathogens. Nature, 2010, 468: 527-532.

[23]Wu J L, Wu C J, Lei C L, Baraodidan M, Bordeos A, Madamba R S, Ramos-Pamplona M, Mauleon R, Portugal A, Ulat V J, Bruskiewich R, Wang G L, Leachi J, Khush G, Leung H. Chmical-and irradiation-induced mutants of indica rice IR64 for forward and reverse genetics. Plant Molecular Biology, 2005, 59: 85-97.

[24]Lin X H, Zhang D P, Xie Y F, Gao H P, Zhang Q. Identification and mapping of a new gne for bacterial blight resistance in rice markers. Phytopathology, 1996, 86: 1156-1159.

[25]Misteli T, Spector D L. Applications of the green fluorescent protein in cell biology and biotechnology. Nature Biotechnology, 1997, 15(10): 961-964.

[26]Yeh E, Gustafson K, Boulianne G L. Green fluorescent protein as a vital marker and reporter of gene expression in Drosophila. Proceedings of the National Academy of Sciences of the USA, 1995, 92: 7036-7040.

[27]Jen S, Seongbin H, Yasuo N, Hirokazu K, David W. Green-fluorescent protein as a new vital marker in plant cells. The Plant Journal, 1995, 8(5): 777-784.

[28]殷晓敏, 徐碧玉, 郑  雯, 曾会才, 马蔚红, 王家保, 李焕苓, 金志强. 香蕉枯萎病菌侵染香蕉根系的组织学过程. 植物病理学报, 2011, 41(6): 570-575.

Yin X M, Xu B Y, Zheng W, Zeng H C, Ma W H, Wang J B, Li H L, Jin Z Q. Histological observation of banana root infected by Fusarium oxysporum f. sp. cubense. Acta Phytopathologica Sinica, 2011, 41(6): 570-575. (in Chinese)
[1] PENG TingShen, LU JiuYan, WU MeiLin, YAN YuXin, LIU HongZhou, NAN WenBin, QIN XiaoJian, LI Ming, GONG JunYi, LIANG YongShu. QTL Analysis of Yield-Related Traits in Both Huangnuo2# and Changbai7# of Perennial Chinese Rice [J]. Scientia Agricultura Sinica, 2026, 59(7): 1361-1379.
[2] CUI JieHao, ZHANG Meng, WANG Qin, YU JiaYan, LIN Kun, LI ShangZe, LAN Heng, GENG YanQiu, ZHANG Qiang, GUO LiYing, SHAO XiWen. Evaluation of Lodging Resistance and Its Physiological Mechanisms in Japonica Rice Resources [J]. Scientia Agricultura Sinica, 2026, 59(7): 1420-1438.
[3] YUAN HaoLiang, NIE Jun, LI Peng, LU YanHong, LIAO YuLin, XU ChangXu, LI ZhongYi, CAO WeiDong, ZHANG JiangLin. Effects of Co-Utilization of Chinese Milk Vetch and Rice Straw on Soil Phosphorus Composition and Phosphorus Activation of Paddy Field in Southern China [J]. Scientia Agricultura Sinica, 2026, 59(7): 1480-1491.
[4] XU YangHaoJun, CHEN LiMing, YANG ShiQi, TANG YiFan, TAN XueMing, ZENG YongJun, PAN XiaoHua, ZENG YanHua. Effects of Long-Term Different Straw Returning Methods on Soil Organic Carbon, Nutrients and Aggregate Formation in Different Soil Layers of Double Cropping Rice Field [J]. Scientia Agricultura Sinica, 2026, 59(7): 1492-1506.
[5] LI XingYu, HUANG Rong, XIAN YiMing, TIAN JiaoJiao, MA XiaoJin, YANG QiaoXi, LI Bing, WANG ChangQuan. Characteristics of Organic Carbon Fractions and Carbon Dioxide Emissions of Different Size Aggregates in Rice Field Soils in Response to Long-Term Fertilization [J]. Scientia Agricultura Sinica, 2026, 59(6): 1255-1271.
[6] MA ZhaoHui, QUAN ChengZhe, CHENG HaiTao, YANG KanJie, LI XinRui, LÜ WenYan. The Breeding Goals and Strategies of Northeast Japonica Rice Under the Background of Zhongke Fa No.5 [J]. Scientia Agricultura Sinica, 2026, 59(5): 927-936.
[7] ZHANG WeiJian, YAN ShengJi, SHANG ZiYin, TANG ZhiWei, WU LiuGe, LI JiaRui, CHEN HaoTian, DENG AiXing, ZHANG Jun, ZHANG Xin, ZHENG ChengYan, SONG ZhenWei. Methane Emissions from Paddy Fields: Not Entirely Attributable to Rice Cultivation [J]. Scientia Agricultura Sinica, 2026, 59(4): 824-833.
[8] CHEN Min, JIAO ZiLan, QIAO ChengBin, XU Hao, ZHANG Bi, MA DongHua, KONG WeiRu, WANG JingWen, SONG JiaWei, LUO ChengKe, LI PeiFu, TIAN Lei. Morpho-Physiological Responses and Adaptive Strategies of Rice Germplasm Accessions from Different Subspecies Under Salt Stress [J]. Scientia Agricultura Sinica, 2026, 59(4): 705-722.
[9] GUO FuCheng, TANG HaiJiang, HAO XinYi, MA GuoLin, YANG JiuJu, HUANG LinFeng, TIAN Lei, WANG Bin, LUO ChengKe. Effects of Different Irrigation Methods on Water-Salt Transport, Rice Yield, and Water Use Efficiency in Saline Soil in Ningxia [J]. Scientia Agricultura Sinica, 2026, 59(4): 750-764.
[10] LUO Wei, YU Hong, YUAN LiXin, WANG LingLing, ZHAO JinPeng, YIN Wei, WANG MingTian, WANG RuLin. Change of Geographic Distributions of Ratoon Rice in Sichuan- Chongqing Under Global Climate Change [J]. Scientia Agricultura Sinica, 2026, 59(4): 765-780.
[11] ZHU Shu, GUO ZhiPeng, SUN Ying. Functional Analysis of Rice Target of Rapamycin OsTOR in Regulating Root Elongation [J]. Scientia Agricultura Sinica, 2026, 59(3): 475-485.
[12] LÜ WenYan, CHENG HaiTao, MA ZhaoHui, TIAN ShuHua. Discussion on Hybridization Breeding Technology and Strategy of Rice in the New Era of Breeding [J]. Scientia Agricultura Sinica, 2026, 59(2): 233-238.
[13] LIAO TingLu, SHI YaFei, XIAO DongHao, SHE YangMengFei, GUO FuCheng, YANG JiuJu, TANG HaiJiang, LUO ChengKe. The Effect of Exogenous Nitroprusside on Sugar Metabolism in Rice Seedlings Under Alkaline Stress [J]. Scientia Agricultura Sinica, 2026, 59(2): 265-277.
[14] LIU TianSheng, LIU GengYuan, ZHAO AnQi, YANG Xu, CAI MingXue, YANG AiWen, LOU MingXuan, LI MuKai, WANG Han, ZHANG YaLing. Pathogenic Population of Rice Bakanae Disease in Heilongjiang Province [J]. Scientia Agricultura Sinica, 2026, 59(2): 305-321.
[15] WANG ZhongNi, LEI Yue, LI JiaLi, GONG YanLong, ZHU SuSong. Functions of ABC Transporter OsARG1 in Rice Heading Date Regulation [J]. Scientia Agricultura Sinica, 2026, 59(1): 1-16.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!