Cloning and characterization of a tau glutathione S-transferase subunit encoding gene in Gossypium hirsutum

Li, Zhikun; Wang, Xingfen; Ma, Jun; Zhang, Guiyin; Ma, Zhiying

doi:10.1266/ggs.83.219

Cited by 5 publications

(5 citation statements)

References 28 publications

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“…ND601. Furthermore, the Gh_A09G1509 ( GhGST ) product exhibited obvious activity against 1‐chloro‐2,4‐dinitrobenzene (CDNB) substrate in our previous study (Li et al ., ). These results demonstrate that the GST genes related to Verticillium wilt resistance in the A‐subgenome are almost twice as abundant as those found in the D‐subgenome, which is rather different from the findings obtained for genes associated with stress tolerance, which are mainly located in the D‐subgenome (Zhang et al ., ).…”

Section: Resultsmentioning

confidence: 97%

“…Specifically, after infection with V. dahliae , the root xylem vessels are occluded, leading to yield reduction and a decrease in fiber quality (Sink and Grey, ; Zhang et al ., ). In a previous study, we cloned the tau‐class gene GhGST (EU074792) from the SSH library under V. dahliae stress (Li et al ., ), and found that the transcriptional levels of glutathione S ‐transferase (GST, E.C.2.5.1.18) genes could also be significantly induced in G. barbadense under V. dahliae stress (Zhang et al ., ). These indicated that GST genes are involved in cotton resistance to Verticillium wilt.…”

Section: Introductionmentioning

confidence: 99%

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A newly identified cluster of glutathione S‐transferase genes provides Verticillium wilt resistance in cotton

Chen

et al. 2019

The Plant Journal

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These authors contributed equally to this work. SUMMARYAs the largest cultivated fiber crop in the world, cotton (Gossypium hirsutum) is often exposed to various biotic stresses during its growth periods. Verticillium wilt caused by Verticillium dahliae is a severe disease in cotton, and the molecular mechanism of cotton resistance for Verticillium wilt needs to be further investigated. Here, we revealed that the cotton genome contains nine types of GST genes. An evolutionary analysis showed that a newly identified cluster (including Gh_A09G1508, Gh_A09G1509 and Gh_A09G1510) located on chromosome 09 of the A-subgenome was under positive selection pressure during the formation of an allotetraploid. Transcriptome analysis showed that this cluster participates in Verticillium wilt resistance. Because the Gh_A09G1509 gene showed the greatest differential expression in the resistant cultivar under V. dahliae stress, we overexpressed this gene in tobacco and found that its overexpression resulted in enhanced Verticillium wilt resistance. Suppression of the gene cluster via virus-induced gene silencing made cotton plants of the resistant cultivar Nongda601 significantly susceptible. These results demonstrated that the GST cluster played an important role in Verticillium wilt resistance. Further investigation showed that the encoded enzymes of the cluster were essential for the delicate equilibrium between the production and scavenging of H 2 O 2 during V. dahliae stress.

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Section: Resultsmentioning

confidence: 97%

Section: Introductionmentioning

confidence: 99%

A newly identified cluster of glutathione S‐transferase genes provides Verticillium wilt resistance in cotton

Chen

et al. 2019

The Plant Journal

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“…These ESTs were assembled into 23,126 unigenes. Annotation results showed that this library contained many previously reported key response genes, such as for PR protein, chitinase, members of the GST gene family, PAL, CYP, and NDR1 (McFadden et al, [63]; Li et al, [64]; Gao et al, [22]; Xu et al, [12]; Zhu et al, [65]; Ahmed et al, [66]). Our library also included several genes that function in the development of cotton fibers, e.g., SusA1 , CesAs , UXS1 , ADF1 , tubulin , and aquaporin (Pan et al, [56]; Yuan et al, [48]; Jiang et al, [67]; Kim et al, [68]; Chi et al, [54]).…”

Section: Discussionmentioning

confidence: 99%

Transcriptome profiling of Gossypium barbadense inoculated with Verticillium dahliae provides a resource for cotton improvement

et al. 2013

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BackgroundVerticillium wilt, caused by the fungal pathogen Verticillium dahliae, is the most severe disease in cotton (Gossypium spp.), causing great lint losses worldwide. Disease management could be achieved in the field if genetically improved, resistant plants were used. However, the interaction between V. dahliae and cotton is a complicated process, and its molecular mechanism remains obscure. To understand better the defense response to this pathogen as a means for obtaining more tolerant cultivars, we monitored the transcriptome profiles of roots from resistant plants of G. barbadense cv. Pima90-53 that were challenged with V. dahliae.ResultsIn all, 46,192 high-quality expressed sequence tags (ESTs) were generated from a full-length cDNA library of G. barbadense. They were clustered and assembled into 23126 unigenes that comprised 2661 contigs and 20465 singletons. Those unigenes were assigned Gene Ontology terms and mapped to 289 KEGG pathways. A total of 3027 unigenes were found to be homologous to known defense-related genes in other plants. They were assigned to the functional classification of plant–pathogen interactions, including disease defenses and signal transduction. The branch of "SA→NPR1→TGA→PR-1→Disease resistance" was first discovered in the interaction of cotton–V. dahliae, indicating that this wilt process includes both biotrophic and necrotrophic stages. In all, 4936 genes coding for putative transcription factors (TF) were identified in our library. The most abundant TF family was the NAC group (527), followed by G2-like (440), MYB (372), BHLH (331), bZIP (271) ERF, C3H, and WRKY. We also analyzed the expression of genes involved in pathogen-associated molecular pattern (PAMP) recognition, the activation of effector-triggered immunity, TFs, and hormone biosynthesis, as well as genes that are pathogenesis-related, or have roles in signaling/regulatory functions and cell wall modification. Their differential expression patterns were compared among mock-/inoculated- and resistant/susceptible cotton. Our results suggest that the cotton defense response has significant transcriptional complexity and that large accumulations of defense-related transcripts may contribute to V. dahliae resistance in cotton. Therefore, these data provide a resource for cotton improvement through molecular breeding approaches.ConclusionsThis study generated a substantial amount of cotton transcript sequences that are related to defense responses against V. dahliae. These genomics resources and knowledge of important related genes contribute to our understanding of host–pathogen interactions and the defense mechanisms utilized by G. barbadense, a non-model plant system. These tools can be applied in establishing a modern breeding program that uses marker-assisted selections and oligonucleotide arrays to identify candidate genes that can be linked to valuable agronomic traits in cotton, including disease resistance.

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“…Only sequences that showed similarity higher than 1e-75 (E-value) were considered as putative homologous to the Arabidopsis genes and were selected for primer design. We also selected the gene encoding the poly-ubiquitin, GhUBQ14 , commonly used in cotton for experiments of Northern blots and RT-qPCRs [26,27] (Table 1). Primers were designed with Primer 3 software [28] using as criterion amplified products from 80 to 180 bp with a Tm of 60 ± 1°C (primer sequences are shown in Table 1).…”

Section: Methodsmentioning

confidence: 99%

Identification and evaluation of new reference genes in Gossypium hirsutumfor accurate normalization of real-time quantitative RT-PCR data

et al. 2010

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BackgroundNormalizing through reference genes, or housekeeping genes, can make more accurate and reliable results from reverse transcription real-time quantitative polymerase chain reaction (qPCR). Recent studies have shown that no single housekeeping gene is universal for all experiments. Thus, suitable reference genes should be the first step of any qPCR analysis. Only a few studies on the identification of housekeeping gene have been carried on plants. Therefore qPCR studies on important crops such as cotton has been hampered by the lack of suitable reference genes.ResultsBy the use of two distinct algorithms, implemented by geNorm and NormFinder, we have assessed the gene expression of nine candidate reference genes in cotton: GhACT4, GhEF1α5, GhFBX6, GhPP2A1, GhMZA, GhPTB, GhGAPC2, GhβTUB3 and GhUBQ14. The candidate reference genes were evaluated in 23 experimental samples consisting of six distinct plant organs, eight stages of flower development, four stages of fruit development and in flower verticils. The expression of GhPP2A1 and GhUBQ14 genes were the most stable across all samples and also when distinct plants organs are examined. GhACT4 and GhUBQ14 present more stable expression during flower development, GhACT4 and GhFBX6 in the floral verticils and GhMZA and GhPTB during fruit development. Our analysis provided the most suitable combination of reference genes for each experimental set tested as internal control for reliable qPCR data normalization. In addition, to illustrate the use of cotton reference genes we checked the expression of two cotton MADS-box genes in distinct plant and floral organs and also during flower development.ConclusionWe have tested the expression stabilities of nine candidate genes in a set of 23 tissue samples from cotton plants divided into five different experimental sets. As a result of this evaluation, we recommend the use of GhUBQ14 and GhPP2A1 housekeeping genes as superior references for normalization of gene expression measures in different cotton plant organs; GhACT4 and GhUBQ14 for flower development, GhACT4 and GhFBX6 for the floral organs and GhMZA and GhPTB for fruit development. We also provide the primer sequences whose performance in qPCR experiments is demonstrated. These genes will enable more accurate and reliable normalization of qPCR results for gene expression studies in this important crop, the major source of natural fiber and also an important source of edible oil. The use of bona fide reference genes allowed a detailed and accurate characterization of the temporal and spatial expression pattern of two MADS-box genes in cotton.

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Cloning and characterization of a tau glutathione S-transferase subunit encoding gene in Gossypium hirsutum

Cited by 5 publications

References 28 publications

A newly identified cluster of glutathione S‐transferase genes provides Verticillium wilt resistance in cotton

A newly identified cluster of glutathione S‐transferase genes provides Verticillium wilt resistance in cotton

Transcriptome profiling of Gossypium barbadense inoculated with Verticillium dahliae provides a resource for cotton improvement

Identification and evaluation of new reference genes in Gossypium hirsutumfor accurate normalization of real-time quantitative RT-PCR data

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