Welcome to Acta Agronomica Sinica，

Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (05): 764-771.doi: 10.3724/SP.J.1006.2011.00764

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

Isolation of NBS-LRR Late Blight Resistance Genes from Potato BAC Library by System of Magnetic Separation

XU Jian-Fei, JIN Li-Ping, PANG Wan-Fu, BIAN Chun-Song, DUAN Shao-Guang, LIU Jie, HUANG San-Wen, and QU Dong-Yu*   

  1. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2010-11-18 Revised:2011-03-08 Online:2011-05-12 Published:2011-03-24
  • Contact: 屈冬玉, E-mail: dyqu@mail.caas.net.cn, Tel: 010-82105943

PDF

1956

Abstract: Cultivated potato (Solanum tuberosum) is an auto-tetraploid crop, which is very difficult to isolate late blight resistance genes. Nevertheless, it is discovered that there is conseved NBS-LRR domain among late blight resistance genes. In this study, conserved probes of R3a late blight resistance genes family were developed by alignment of sequences. And then, the DNA of BAC SH23G23 containingthe R3a gene was partially digested into 7–11 kb fragments. By magnetic separation system combined with conserved probes, the 7–11 kb fragments were enriched and cloned into binary vector pBINPLUS. Through identification of positive clones and colony PCR, the ratio of clones including R3a gene to all positive clones reached 82.76%, which was nearly 19 times higher than enrichment before. The system of magnetic separation for R genes and their analogs established in this study provides a new strategy for conserved domain genes cloning from polyploid crops.

Key words: Potato, Late blight, Resistance genes, Magnetic separation

[1]Meyer R C, Milbourne D, Hackett C A, Bradshaw J E, McNichol J W, Waugh, R. Linkage analysis in tetraploid potato and association of markers with quantitative resistance to late blight (Phytophthora infestans). Mol Gen Genet, 1998, 259: 150–160
[2]Dangl J L, Jones J D. Plant pathogens and integrated defence responses to infection. Nature, 2001, 411: 826–833
[3]Ballvora A, Ercolano M R, Weiss J, Meksem K, Bormann C A, Oberhagemann P, Salamini F, Gebhardt C. The R1 gene for potato resistance to late blight (Phytophthora infestans) belongs to the leucine zipper/NBS/LRR class of plant resistance genes. Plant J, 2002, 30: 361–371
[4]Song J, Bradeen J M, Naess S K, Raasch J A, Wielgus S M, Haberlach G T, Liu J, Kuang H, Austin-Phillips S, Buell C R, Helgeson J P, Jiang J. Gene RB cloned from Solanum bulbocastanum confers broad spectrum resistance to potato late blight. Proc Nat Acad Sci USA, 2003, 100: 9128–9133
[5]van der Vossen E A, Sikkema A, Hekkert B L, Gros J, Stevens P, Muskens M, Wouters D, Pereira A, Stiekema W, Allefs S. An ancient R gene from the wild potato species Solanum bulbocastanum confers broad-spectrum resistance to Phytophthora infestans in cultivated potato and tomato. Plant J, 2003, 36: 867–882
[6]Huang S, van der Vossen E A, Kuang H H, Vleeshouwers V G A A, Zhang N W, Borm T J A, van Eck H J, Baker B, Jacobsen E, Visser R G F. Comparative genomics enabled the isolation of the R3a late blight resistance gene in potato. Plant J, 2005, 42: 251–261
[7]van der Vossen E A, Gros J, Sikkema A, Muskens M, Wouters D, Wolters P, Pereira A, Allefs S. The Rpi-blb2 gene from Solanum bulbocastanum is an Mi-1 gene homolog conferring broad-spectrum late blight resistance in potato. Plant J, 2005, 44: 208–222
[8]Huang S. Discovery and characterization of the major late blight resistance complex in potato: genomic structure, functional diversity, and implications. the Netherlands: Wageningen University, 2005
[9]Sambrook J, Russell D W. Molecular Cloning: a Laboratory Manual, 3rd ed. New York: Cold Spring Harbor Laboratory Press, 2001
[10]Coulson A, Sulston J, Brenner S, Karn J. Toward a physical map of the genome of the nematode Caenorhabditis elegans. ProeNatn Acad Sci USA, 1986, 83: 7821–7825
[11]Shiu S, Karlowski W M, Pan R, Tzeng Y, Mayer K F X, Lia W. Comparative analysis of the receptor-like kinase family in arabidopsis and rice. Plant Cell, 2004, 16: 1220–1234
[12]Leister D, Ballvora A, Salamini F, Gebhardt C. A PCR based approach for isolating pathogen resistance genes from potato with potential for wide application in plants. Nat Genet, 1996, 14: 421–429
[13]Bender K, Schneider P M. Validation and casework testing of the BioPlex211 for STR typing of telogen hair roots. Forensic Sci Int, 2006, 161: 52–59
[14]Li D(李炟), Ji K-Z(季孔庶), Gong J(龚佳). Microsatellite enrichment by magnetic beads in Pinus massoniana. Mol Plant Breed(分子植物育种), 2007, 5(1): 141–144 (in Chinese with English abstract)
[1] WANG Hai-Bo, YING Jing-Wen, HE Li, YE Wen-Xuan, TU Wei, CAI Xing-Kui, SONG Bo-Tao, LIU Jun. Identification of chromosome loss and rearrangement in potato and eggplant somatic hybrids by rDNA and telomere repeats [J]. Acta Agronomica Sinica, 2022, 48(5): 1273-1278.
[2] SHI Yan-Yan, MA Zhi-Hua, WU Chun-Hua, ZHOU Yong-Jin, LI Rong. Effects of ridge tillage with film mulching in furrow on photosynthetic characteristics of potato and yield formation in dryland farming [J]. Acta Agronomica Sinica, 2022, 48(5): 1288-1297.
[3] FENG Ya, ZHU Xi, LUO Hong-Yu, LI Shi-Gui, ZHANG Ning, SI Huai-Jun. Functional analysis of StMAPK4 in response to low temperature stress in potato [J]. Acta Agronomica Sinica, 2022, 48(4): 896-907.
[4] ZHANG Xia, YU Zhuo, JIN Xing-Hong, YU Xiao-Xia, LI Jing-Wei, LI Jia-Qi. Development and characterization analysis of potato SSR primers and the amplification research in colored potato materials [J]. Acta Agronomica Sinica, 2022, 48(4): 920-929.
[5] JIN Rong, JIANG Wei, LIU Ming, ZHAO Peng, ZHANG Qiang-Qiang, LI Tie-Xin, WANG Dan-Feng, FAN Wen-Jing, ZHANG Ai-Jun, TANG Zhong-Hou. Genome-wide characterization and expression analysis of Dof family genes in sweetpotato [J]. Acta Agronomica Sinica, 2022, 48(3): 608-623.
[6] TAN Xue-Lian, GUO Tian-Wen, HU Xin-Yuan, ZHANG Ping-Liang, ZENG Jun, LIU Xiao-Wei. Characteristics of microbial community in the rhizosphere soil of continuous potato cropping in arid regions of the Loess Plateau [J]. Acta Agronomica Sinica, 2022, 48(3): 682-694.
[7] ZHANG Hai-Yan, XIE Bei-Tao, JIANG Chang-Song, FENG Xiang-Yang, ZHANG Qiao, DONG Shun-Xu, WANG Bao-Qing, ZHANG Li-Ming, QIN Zhen, DUAN Wen-Xue. Screening of leaf physiological characteristics and drought-tolerant indexes of sweetpotato cultivars with drought resistance [J]. Acta Agronomica Sinica, 2022, 48(2): 518-528.
[8] JIAN Hong-Ju, SHANG Li-Na, JIN Zhong-Hui, DING Yi, LI Yan, WANG Ji-Chun, HU Bai-Geng, Vadim Khassanov, LYU Dian-Qiu. Genome-wide identification and characterization of PIF genes and their response to high temperature stress in potato [J]. Acta Agronomica Sinica, 2022, 48(1): 86-98.
[9] XU De-Rong, SUN Chao, BI Zhen-Zhen, QIN Tian-Yuan, WANG Yi-Hao, LI Cheng-Ju, FAN You-Fang, LIU Yin-Du, ZHANG Jun-Lian, BAI Jiang-Ping. Identification of StDRO1 gene polymorphism and association analysis with root traits in potato [J]. Acta Agronomica Sinica, 2022, 48(1): 76-85.
[10] ZHANG Si-Meng, NI Wen-Rong, LYU Zun-Fu, LIN Yan, LIN Li-Zhuo, ZHONG Zi-Yu, CUI Peng, LU Guo-Quan. Identification and index screening of soft rot resistance at harvest stage in sweetpotato [J]. Acta Agronomica Sinica, 2021, 47(8): 1450-1459.
[11] SONG Tian-Xiao, LIU Yi, RAO Li-Ping, Soviguidi Deka Reine Judesse, ZHU Guo-Peng, YANG Xin-Sun. Identification and expression analysis of cell wall invertase IbCWIN gene family members in sweet potato [J]. Acta Agronomica Sinica, 2021, 47(7): 1297-1308.
[12] TANG Rui-Min, JIA Xiao-Yun, ZHU Wen-Jiao, YIN Jing-Ming, YANG Qing. Cloning of potato heat shock transcription factor StHsfA3 gene and its functional analysis in heat tolerance [J]. Acta Agronomica Sinica, 2021, 47(4): 672-683.
[13] LI Peng-Cheng, BI Zhen-Zhen, SUN Chao, QIN Tian-Yuan, LIANG Wen-Jun, WANG Yi-Hao, XU De-Rong, LIU Yu-Hui, ZHANG Jun-Lian, BAI Jiang-Ping. Key genes mining of DNA methylation involved in regulating drought stress response in potato [J]. Acta Agronomica Sinica, 2021, 47(4): 599-612.
[14] QIN Tian-Yuan, LIU Yu-Hui, SUN Chao, BI Zhen-Zhen, LI An-Yi, XU De-Rong, WANG Yi-Hao, ZHANG Jun-Lian, BAI Jiang-Ping. Identification of StIgt gene family and expression profile analysis of response to drought stress in potato [J]. Acta Agronomica Sinica, 2021, 47(4): 780-786.
[15] JIANG Wei, PAN Zhe-Chao, BAO Li-Xian, ZHOU Fu-Xian, LI Yan-Shan, SUI Qi-Jun, LI Xian-Ping. Genome-wide association analysis for late blight resistance of potato resources [J]. Acta Agronomica Sinica, 2021, 47(2): 245-261.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd