Differentially Expressed Genes of Soybean During Infection by Phytophthorasojae

doi:10.1016/S1671-2927(00)8554

Journal of Integrative Agriculture ›› 2012, Vol. 12 ›› Issue (3): 368-377.DOI: 10.1016/S1671-2927(00)8554

Differentially Expressed Genes of Soybean During Infection by Phytophthorasojae

XU Peng-fei, Allen Xue, CHEN Wei-yuan, LV Hui-ying, FAN Su-jie, WANG Xin, JIANG Liang-yu, ZHANG Shu-zhen

1.Key Laboratory of Soybean Biology, Ministry of Education/Soybean Research Institute, Northeast Agricultural University, Harbin
150030, P.R.China
2.Soybean Research Institute, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, P.R.China
3.Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, Ontario K1A 0C6, Canada
4.Suihua Research Institute, Heilongjiang Academy of Agricultural Sciences, Suihua 152052, P.R.China
5.Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, P.R.China
6.College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, P.R.China

收稿日期:2011-01-18 出版日期:2012-03-01 发布日期:2012-03-11
通讯作者: Correspondence ZHANG Shu-zhen, Tel: +86-451-55191487, Fax: +86-451-55190447, E-mail: dnzhangshuzhen@yahoo.com
基金资助:
This research was supported by the Program for New CenturyExcellent Talents in Universities, Ministry of Education,China (NCET-09-164), the National Natural Science Foundationof China (30671317, 30971811, 31071439, and31110103001), the Program for New Century Excellent Talentsin Universities in Heilongjiang Province, China (NCET-06-007), the Natural Science Foundation of HeilongjiangProvince, China (C200814).

Differentially Expressed Genes of Soybean During Infection by Phytophthorasojae

XU Peng-fei, Allen Xue, CHEN Wei-yuan, LV Hui-ying, FAN Su-jie, WANG Xin, JIANG Liang-yu, ZHANG Shu-zhen

1.Key Laboratory of Soybean Biology, Ministry of Education/Soybean Research Institute, Northeast Agricultural University, Harbin
150030, P.R.China
2.Soybean Research Institute, Heilongjiang Academy of Agricultural Sciences, Harbin 150086, P.R.China
3.Eastern Cereal and Oilseed Research Centre, Agriculture and Agri-Food Canada, Ontario K1A 0C6, Canada
4.Suihua Research Institute, Heilongjiang Academy of Agricultural Sciences, Suihua 152052, P.R.China
5.Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, P.R.China
6.College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, P.R.China

Received:2011-01-18 Online:2012-03-01 Published:2012-03-11
Contact: Correspondence ZHANG Shu-zhen, Tel: +86-451-55191487, Fax: +86-451-55190447, E-mail: dnzhangshuzhen@yahoo.com
Supported by:
This research was supported by the Program for New CenturyExcellent Talents in Universities, Ministry of Education,China (NCET-09-164), the National Natural Science Foundationof China (30671317, 30971811, 31071439, and31110103001), the Program for New Century Excellent Talentsin Universities in Heilongjiang Province, China (NCET-06-007), the Natural Science Foundation of HeilongjiangProvince, China (C200814).

摘要/Abstract

摘要： To elucidate the differential gene expression patterns in soybeans during infection by Phytophthora sojae, a cDNA libraryfor suppression subtractive hybridization (SSH) was constructed with cDNAs from soybean cultivar Suinong 10 treatedwith sterile distilled water as the driver and cDNAs from Suinong 10 inoculated with P. sojae as the tester. A total of 2 067recombinant colonies from the SSH library were randomly picked, amplified, and sequenced. After discarding 312 poorquality expressed sequence tags (EST), 1 755 high quality ESTs were assembled and edited to 1 384 tentatively uniquegenes (TUG), in which, 586 showed significant homology to known sequences, and 798 had low homology or no matchwith the known sequences. A cDNA microarray containing 307 singletons from the 586 TUGs and 222 singletons from the798 TUGs was developed to characterize differentially expressed cDNAs in the SSH library, and eight cDNAs wereidentified to be up-regulated after microarray analysis and then confirmed by real-time PCR. They were homologous to theprotein 10, and were also related to some proteins in disease resistance response, such as pathogen-related protein,phenylalanine ammonia-lyase, isoflavone reductase, WRKY transcription factor 31, major allergen Pru ar 1, and pleiotropicdrug resistance protein 12. Most of the up-regulated cDNAs encode enzymes of phytoalexin biosynthesis andpathogenesis-related proteins involved in plant disease resistance. Here, we fist reported the Pru ar 1 in soybeans. Thefindings of this research have contributed to better understanding of soybean resistance to P. sojae at the molecular level.

关键词: cDNA microarray, Glycine max, Phytophthora sojae, soybean, suppressed subtraction hybridization

Abstract: To elucidate the differential gene expression patterns in soybeans during infection by Phytophthora sojae, a cDNA libraryfor suppression subtractive hybridization (SSH) was constructed with cDNAs from soybean cultivar Suinong 10 treatedwith sterile distilled water as the driver and cDNAs from Suinong 10 inoculated with P. sojae as the tester. A total of 2 067recombinant colonies from the SSH library were randomly picked, amplified, and sequenced. After discarding 312 poorquality expressed sequence tags (EST), 1 755 high quality ESTs were assembled and edited to 1 384 tentatively uniquegenes (TUG), in which, 586 showed significant homology to known sequences, and 798 had low homology or no matchwith the known sequences. A cDNA microarray containing 307 singletons from the 586 TUGs and 222 singletons from the798 TUGs was developed to characterize differentially expressed cDNAs in the SSH library, and eight cDNAs wereidentified to be up-regulated after microarray analysis and then confirmed by real-time PCR. They were homologous to theprotein 10, and were also related to some proteins in disease resistance response, such as pathogen-related protein,phenylalanine ammonia-lyase, isoflavone reductase, WRKY transcription factor 31, major allergen Pru ar 1, and pleiotropicdrug resistance protein 12. Most of the up-regulated cDNAs encode enzymes of phytoalexin biosynthesis andpathogenesis-related proteins involved in plant disease resistance. Here, we fist reported the Pru ar 1 in soybeans. Thefindings of this research have contributed to better understanding of soybean resistance to P. sojae at the molecular level.

Key words: cDNA microarray, Glycine max, Phytophthora sojae, soybean, suppressed subtraction hybridization

XU Peng-fei, Allen Xue, CHEN Wei-yuan, LV Hui-ying, FAN Su-jie, WANG Xin, JIANG Liang-yu, ZHANG Shu-zhen. Differentially Expressed Genes of Soybean During Infection by Phytophthorasojae[J]. Journal of Integrative Agriculture, 2012, 12(3): 368-377.

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Differentially Expressed Genes of Soybean During Infection by Phytophthorasojae

Differentially Expressed Genes of Soybean During Infection by Phytophthorasojae

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