Functional Characterization of Arabidopsis thaliana WRKY39 in Heat Stress

Li, Shujia,Zhou, Xiang,Chen, Ligang,Huang, Weidong,Yu, Diqiu Korean Society for Molecular and Cellular Biology 2010 Molecules and cells Vol.29 No.5

Arabidopsis thaliana WRKY39, a transcription factor that is induced by heat stress, is a member of the group II WRKY proteins and responds to both abiotic and biotic stress. Heat-treated seeds and plants of WRKY39 knock-down mutants had increased susceptibility to heat stress, showing reduced germination, decreased survival, and elevated electrolyte leakage compared with wild-type plants. In contrast, WRKY39 over-expressing plants exhibited enhanced thermotolerance compared with wild-type plants. RT-PCR and qRT-PCR analysis of wrky39 mutants and WRKY39 over-expressing plants identified putative genes regulated by WRKY39. Consistent with a role for WRKY39 in heat tolerance, the expression levels of salicylic acid (SA)-regulated PR1 and SA-related MBF1c genes were down-regulated in wrky39 mutants. In contrast, over-expression of WRKY39 increased the expression of PR1 and MBF1c. The WRKY39 transcript was induced in response to treatment with SA or methyljasmonate. Analysis of heat stress-induced WRKY39 in defense signaling mutants, including coi1, ein2, and sid2, further indicated that WRKY39 was positively co-regulated by the SA and jasmonate (JA) signaling pathways. Together, these findings reveal that heat stress-induced WRKY39 positively regulates the cooperation between the SA- and JA-activated signaling pathways that mediate responses to heat stress.

Functional Characterization of Arabidopsis thaliana WRKY39 in Heat Stress

Shujia Li,Xiang Zhou,Ligang Chen,Weidong Huang,Diqiu Yu 한국분자세포생물학회 2010 Molecules and cells Vol.29 No.5

Arabidopsis thaliana WRKY39, a transcription factor that is induced by heat stress, is a member of the group II WRKY proteins and responds to both abiotic and biotic stress. Heat-treated seeds and plants of WRKY39 knock-down mutants had increased susceptibility to heat stress, show-ing reduced germination, decreased survival, and elevated electrolyte leakage compared with wild-type plants. In con-trast, WRKY39 over-expressing plants exhibited enhanced thermotolerance compared with wild-type plants. RT-PCR and qRT-PCR analysis of wrky39 mutants and WRKY39 over-expressing plants identified putative genes regulated by WRKY39. Consistent with a role for WRKY39 in heat tolerance, the expression levels of salicylic acid (SA)-regulated PR1 and SA-related MBF1c genes were down-regulated in wrky39 mutants. In contrast, over-expression of WRKY39 increased the expression of PR1 and MBF1c. The WRKY39 transcript was induced in response to treat-ment with SA or methyljasmonate. Analysis of heat stress-induced WRKY39 in defense signaling mutants, including coi1, ein2, and sid2, further indicated that WRKY39 was positively co-regulated by the SA and jasmonate (JA) sig-naling pathways. Together, these findings reveal that heat stress-induced WRKY39 positively regulates the coopera-tion between the SA- and JA-activated signaling pathways that mediate responses to heat stress.

억새 EST 정보 유래 전사요소 WRKY의 난지형 잔디의 저온 발현 반응성

정성진,최영인,이긍주 한국잔디학회 2013 Weed & Turfgrass Science Vol.2 No.4

Whole genome transcriptomes from Miscanthus species were sequenced and analyzed, which provided 50 different types of transcription factor (TF) involving various developmental processes or environmental stresses. Among the explored TF, WRKY gene family was the major type and one of the WRKY genes, MSIR7180_WRKY4, induced under low temperature environment was selected to investigate how the Miscanthus- originated MSIR7180_WRKY4 TF responds when exposed to low temperature in four warm-season turfgrasses (Z.matrella ‘Semil’, bermudagrass, St. Augustinegrass, and seashore paspalum). The MSIR7180_WRKY4 was expressed higher during low temperature period in Bermudagrass, but the expression was enhanced in St. Augustinegrass. In contrast, the gene in ‘Semil’ cultivar was barely expressed and relatively less expressed, but repressed gradually in seashore paspalum, which seems to allow two turfgrasses stay-green longer in the fall season. The results indicate that bermudagrass and St. Augustinegrass adapt to low temperature quickly, but relative tolerance to low or cold temperature at the molecular level needs to be further investigated at different physiological stages and the corresponding genes systematically. 국내에 자생하는 참억새(M. sinensis)와 물억새(M.sacchariflorus)의 잎과 지하경 조직 EST로부터 유전자의 전사를 조절하는 전사요소 탐색하여 저온에 반응하는 유전자를 분리하고 난지형 잔디에서 저온 반응의 차이를 알아보기 위하여 본 연구를 실시하였다. 분석 결과 탐색된 전사조절 요소의 종류는 총 50 종류로 나타났고, 그 중 WRKY family에 속하는 EST 절편이 226개로 가장 많이발견되었다(9.6%). 그 중 억새 WRKY family를 기능이 밝혀진 다른 작물의 WRKY 유전자들과 비교 검색한 결과 약80개의 억새 isotig가 저온에 반응하여 발현이 유도 또는 억제되는 것으로 나타났다. 그 중 애기장대와 벼에서 저온 처리 후 발현이 증가되는 것으로 알려진 억새의 MSIR7180_ WRKY4 유전자를 대상으로 그 발현양상을 조사한 결과 버뮤다그래스는 비교구에서와 비슷하게 처리기간 동안 높은 유전자의 발현을 보였고, 금잔디(Z. matrella)간의 교배를 통해 얻어진 세밀 품종은 처리기간 내내 발현이 미약하였다. St. Augustinegrass는 3일 동안은 비교구와 큰 차이가 없다가 5일째부터 발현이 증가하였고, Seashorepaspalum은 처리 초기에 발현이 높다가 처리가 진행되면서 약해지는 경향을 나타냈다. 이 결과로 미루어 볼 때 버뮤다그래스와 St. Augustine grass는 저온 반응성이 신속하여 휴면을 준비하지만 금잔디와 Seashore paspalum은 휴면 돌입이 늦어 녹색이 상대적으로 늦게까지 유지되는 것으로 판단되어 저온 적응을 위한 또 다른 환경요인의 작용이 있을 것으로 여겨진다. 녹색 기간이 길고 내한성이 높은 품종의 개발을 위해서는 더 많은 유전자의 종합적인 고찰과 판단이 요구된다.

Marker Production by PCR Amplification with Primer Pairs from Conserved Sequences of WRKY Genes in Chili Pepper

Kim, Hyounjoung,Lee, Heungryul,Han, Jungheon,Yeom, Seonin,Harn, Cheehark,Kim, Byungdong Korean Society for Molecular Biology 2008 Molecules and cells Vol.25 No.2

Despite increasing awareness of the importance of WRKY genes in plant defense signaling, the locations of these genes in the Capsicum genome have not been established. To develop WRKY-based markers, primer sequences were deduced from the conserved sequences of the DNA binding motif within the WRKY domains of tomato and pepper genes. These primers were derived from upstream and downstream parts of the conserved sequences of the three WRKY groups. Six primer combinations of each WRKY group were tested for polymorphisms between the mapping parents, C. annuum 'CM334' and C. annuum 'Chilsungcho'. DNA fragments amplified by primer pairs deduced from WRKY Group II genes revealed high levels of polymorphism. Using 32 primer pairs to amplify upstream and downstream parts of the WRKY domain of WRKY group II genes, 60 polymorphic bands were detected. Polymorphisms were not detected with primer pairs from downstream parts of WRKY group II genes. Half of these primers were subjected to F2 genotyping to construct a linkage map. Thirty of 41 markers were located evenly spaced on 20 of the 28 linkage groups, without clustering. This linkage map also consisted of 199 AFLP and 26 SSR markers. This WRKY-based marker system is a rapid and simple method for generating sequence-specific markers for plant gene families.

Marker Production by PCR Amplification with Primer Pairs from Conserved Sequences of WRKY Genes in Chili Pepper

Hyoun-Joung Kim,Heung-Ryul Lee,Jung-Heon Han,염선인,Chee-Hark Harn,김병동 한국분자세포생물학회 2008 Molecules and cells Vol.25 No.2

Despite increasing awareness of the importance of WRKY genes in plant defense signaling, the locations of these genes in the Capsicum genome have not been established. To develop WRKY-based markers, primer sequences were deduced from the conserved sequences of the DNA binding motif within the WRKY domains of tomato and pepper genes. These primers were derived from upstream and downstream parts of the conserved sequences of the three WRKY groups. Six primer combinations of each WRKY group were tested for polymorphisms between the mapping parents, C. annuum ‘CM334’ and C. annuum ‘Chilsungcho’. DNA fragments amplified by primer pairs deduced from WRKY Group II genes revealed high levels of polymorphism. Using 32 primer pairs to amplify upstream and downstream parts of the WRKY domain of WRKY group II genes, 60 polymorphic bands were detected. Polymorphisms were not detected with primer pairs from downstream parts of WRKY group II genes. Half of these primers were subjected to F2 genotyping to construct a linkage map. Thirty of 41 markers were located evenly spaced on 20 of the 28 linkage groups, without clustering. This linkage map also consisted of 199 AFLP and 26 SSR markers. This WRKY-based marker system is a rapid and simple method for generating sequence-specific markers for plant gene families.

Biological Network Analyses of WRKY Transcription Factor Family in Soybean (Glycine max) under Low Phosphorus Treatment

Fırat Kurt,Ertugrul Filiz 한국작물학회 2020 Journal of crop science and biotechnology Vol.23 No.2

WRKY transcription factor (TF) is plant specific genes and play essential roles involved in biotic and abiotic stress tolerance. Gene co-expression network (GCN) analysis is effective tool for the interpretation of transcriptomic data. In this study, a co-expression network of 152 WRKY genes using publicly available microarray data (GSE78242) was constructed under low phosphate (Pi) treatment in soybean (Glycine max). A total of 149 nodes and 641 edges were obtained from CGN and seven seed genes were identified. Particularly, Glyma.19G094100 and Glyma.16G054400 seed genes (orthologue to Arabidopsis WRKY75) were found to have a direct connection to P deficiency. Promotor analyses of seed genes revealed the variations in the number of cis-regulatory elements (CREs) ranging from 80 to 137 with a total of 835 CREs. The methylation profile of Glyma.04G218700 (orthologue to Arabidopsis WRKY51) was found higher than other seed genes. As a result, our findings can be used as a scientific basis to cope with P deficiency in soybean as well as abiotic stress tolerance. In addition, these findings of this study may prove the crop improvement studies in future, especially genetically engineered soybean plants.

Identification of a Pathogen-Induced Glycine max Transcription Factor GmWRKY1

Kang, Sang-Gu,Park, Eui-Ho,Do, Kum-Sook The Korean Society of Plant Pathology 2009 Plant Pathology Journal Vol.25 No.4

On screening pathogen-resistant soybean, we identified a WRKY type transcription factor named a Glycine max WRKY1 (GmWRKY1). Expression of GmWRKY1 gene was induced in the soybean sprout by Pseudomonas infection. The GmWRKY1 was expressed in all of the tissues with high levels in stems, leaves and developing seeds. The protein Gm WRKY1 contains highly conserved two WRKY DNA-binding domains having two $C_2-H_2$ zinc-finger motif ($C-X_{4-5}-C-X_{22-23}-H-X-H$) in its N-terminal and C-terminal amino acid sequences. In electrophoresis mobility shift assay, the GmWRKY1 protein bound specifically to W-box elements in the promoters of defense related genes. These results demonstrated that GmWRKY1 is one of the soybean WRKY family genes and the plant-specific transcription factors for defense processes.

The role of WRKY47 gene in regulating selenium tolerance in Arabidopsis thaliana

Xi Wu,Manzhi Tao,Yun Meng,Xiangyu Zhu,Liangwen Qian,Alia Shah,Wei Wang,Shuqing Cao 한국식물생명공학회 2020 Plant biotechnology reports Vol.14 No.1

Selenium stress has a serious impact on the growth of plants. However, the available understanding on the mechanism of Se stress response is still insufficient. In this study, we found that the WRKY47 gene plays an important role in maintaining Se homeostasis and tolerance in Arabidopsis thaliana. Expression of WRKY47 is induced by Se stress. The wrky47 mutants are sensitive to Se stress, and this sensitivity is associated with a decrease in the transcription levels of Se-detoxify genes HMT1 and HMT3. Surprisingly, the WRKY47-overexpression lines also showed increased sensitivity to Se stress and had increased Se content; this correlates to an increase in the transcription level of Se-uptake gene PHT1;4 but not Se-detoxify genes HMT1 and HMT3. Our results suggest that gain- and loss-of-function mutations in WRKY47 enhance the sensitivity of A. thaliana plants to Se stress through different mechanisms.

Deciphering genome-wide WRKY gene family of Triticum aestivum L. and their functional role in response to Abiotic stress

Saurabh Gupta,Vinod Kumar Mishra,Sunita Kumari,Raavi,Ramesh Chand,Pritish Kumar Varadwaj 한국유전학회 2019 Genes & Genomics Vol.41 No.1

WRKY transcription factors (TFs) act in regulating plant growth and development as well as in response to different stress. Some earlier studies done by individual researchers reported different wheat WRKY TFs. Although, the recently released wheat genome has opened an avenue to investigate wheat WRKYs (TaWRKY) TFs. Prime objective of this study to performed genome-wide classifications of TaWRKYs and their functional annotation. The classification of 107 individual identified characterized sequences of TaWRKY (IICS-TaWRKY) and 160 uncharacterized draft sequences of TaWRKY (UDS-TaWRKY), along with their gene structures and motifs analysis was performed. Along with comparative sequence analysis and microarray analysis was performed to mimic out TaWRKYs functions in response to different abiotic stresses, accompanied by in-vitro validation. The comparative phylogenetic analysis and estimation of Ka/Ks ratio with Triticum urartu, illustrate group based clasifications of TaWRKYs and evolutionary divergences. Furthermore, motif-based and protein-DNA interaction analysis of TaWRKYs helps to identify, their putative function in target DNA recognition sites. Subsequently, results of microarray and comparative sequence analysis provides the evidence of TaWRKYs involved in heat and/or drought stress. Further, in-vitro results validates that TaWRKY014, TaWRKY090 are found to participate in response of drought stress, whereas TaWRKY008, TaWRKY122, and WRKY45 are involved in response of heat and drought stress. These findings can be utilized in developing novel heat and drought-tolerant wheat cultivars using marker-assisted breeding and transgenic development.

Identification of a Pathogen-Induced Glycine max Transcription Factor GmWRKY1

강상구,박의호,도금숙 한국식물병리학회 2009 Plant Pathology Journal Vol.25 No.4

On screening pathogen-resistant soybean, we identified a WRKY type transcription factor named a Glycine max WRKY1 (GmWRKY1). Expression of GmWRKY1 gene was induced in the soybean sprout by Pseudomonas infection. The GmWRKY1 was expressed in all of the tissues with high levels in stems, leaves and developing seeds. The protein GmWRKY1 contains highly conserved two WRKY DNA-binding domains having two C2-H2 zinc-finger motif (C-X4-5-C-X22-23-H-XH) in its N-terminal and C-terminal amino acid sequences. In electrophoresis mobility shift assay, the GmWRKY1 protein bound specifically to W-box elements in the promoters of defense related genes. These results demonstrated that GmWRKY1 is one of the soybean WRKY family genes and the plant-specific transcription factors for defense processes.