scholarly journals Introduksi Konstruk Over-Ekspresi Kandidat Gen OsWRKY76 melalui Agrobacterium tumefaciens pada Tanaman Padi Nipponbare

2011 ◽  
Vol 7 (1) ◽  
pp. 19
Author(s):  
Aniversari Apriana ◽  
Atmitri Sisharmini ◽  
Wening Enggarini ◽  
Sudarsono Sudarsono ◽  
Nurul Khumaida ◽  
...  

<p>Delivering of Over-Expression Construct OsWRKY76<br />Candidate Gene in Rice cv. Nipponbare through<br />Agrobacterium tumefaciens. Aniversari Apriana, Atmitri<br />Sisharmini, Wening Enggarini, Sudarsono, Nurul.<br />Khumaida, and Kurniawan R. Trijatmiko. Plant genetic<br />improvement can be done through classical breeding or<br />genetic engineering. WRKY is a transcription factor involved<br />in regulating plant defense responses. OsWRKY76 gene is<br />located in a narrow segment of chromosome 9 which is<br />identified previously to be related to wide spectrum<br />resistance in rice. A sequence of OsWRKY76 (+1.200 bp)<br />has available in the gene bank and it makes possible to<br />isolate, clone, and construct the gene into over-expression<br />vector. The aim of this research was to assemble an overexpression<br />construct of OsWRKY76 candidate gene and<br />introduce it into rice through Agrobacterium-mediated<br />transformation. A construct of pCAMBIA-<br />1301::35S::OsWRKY76 has been successfully assembled and<br />transformed into embryogenic calli of rice cv. Nipponbare<br />using A. tumefaciens strain Agl-1 and EHA 105. A number of<br />126 independent lines has been produced, in which Agl-1<br />showed 3.8 times more efficient than EHA 105. PCR analysis<br />of randomly selected 25 independent lines showed that all<br />of them positively contained hptII gene, a selectable marker<br />used in the over-expression construct of the OsWRKY76<br />candidate gene. Based on the result, it could be concluded<br />that the over-expression construct of OsWRKY76 candidate<br />gene have been successfully introduced into the tissue of<br />Nipponbare.</p>

2021 ◽  
Author(s):  
Honglin Chen ◽  
Liangliang Hu ◽  
Lixia Wang ◽  
Suhua Wang ◽  
Xuzhen Cheng

Abstract Bruchids or seed weevils are serious storage insect pests of mung bean and other pulses. Though bruchid-resistant mung bean germplasm accessions are screened out, the molecular mechanisms of bruchid resistance in mung bean are still unclear. In this study, a segregating population with 182 RILs plants was developed; delimit the controlling gene to a 111-kb physical interval, in which 11 genes were predicted. Vr04g00919 encoding the function of a polygalacturonase inhibitor, was the most likely candidate genes. Here, sequence analysis of the candidate gene coding regions revealed that it has six SNPs between the parental lines and three SNPs resulted in amino acid changes. Sequence alignment revealed that one of these three SNPs are located in a conserved leucine rich repeat (LRR) domain, which is essential for the function of the protein. Subcellular localization of the VrPGIP2-GFP fusion protein indicated that the candidate gene PGIP2 is located in the nucleus and cytosol. RNA-seq and quantitative real-time PCR (qRT-PCR) analysis indicated that many defense responses, cell wall synthesis, biotic and abiotic stresses, and hormone synthesis were greatly activated in the bruchid resistance plants. These findings contribute to the molecular marker assisted selection of bruchid resistance cultivars.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mari Kurokawa ◽  
Masataka Nakano ◽  
Nobutaka Kitahata ◽  
Kazuyuki Kuchitsu ◽  
Toshiki Furuya

AbstractMicroorganisms that activate plant immune responses have attracted considerable attention as potential biocontrol agents in agriculture because they could reduce agrochemical use. However, conventional methods to screen for such microorganisms using whole plants and pathogens are generally laborious and time consuming. Here, we describe a general strategy using cultured plant cells to identify microorganisms that activate plant defense responses based on plant–microbe interactions. Microbial cells were incubated with tobacco BY-2 cells, followed by treatment with cryptogein, a proteinaceous elicitor of tobacco immune responses secreted by an oomycete. Cryptogein-induced production of reactive oxygen species (ROS) in BY-2 cells served as a marker to evaluate the potential of microorganisms to activate plant defense responses. Twenty-nine bacterial strains isolated from the interior of Brassica rapa var. perviridis plants were screened, and 8 strains that enhanced cryptogein-induced ROS production in BY-2 cells were selected. Following application of these strains to the root tip of Arabidopsis seedlings, two strains, Delftia sp. BR1R-2 and Arthrobacter sp. BR2S-6, were found to induce whole-plant resistance to bacterial pathogens (Pseudomonas syringae pv. tomato DC3000 and Pectobacterium carotovora subsp. carotovora NBRC 14082). Pathogen-induced expression of plant defense-related genes (PR-1, PR-5, and PDF1.2) was enhanced by the pretreatment with strain BR1R-2. This cell–cell interaction-based platform is readily applicable to large-scale screening for microorganisms that enhance plant defense responses under various environmental conditions.


2021 ◽  
Vol 22 (8) ◽  
pp. 4214
Author(s):  
Gautam Anand ◽  
Meirav Leibman-Markus ◽  
Dorin Elkabetz ◽  
Maya Bar

Plants lack a circulating adaptive immune system to protect themselves against pathogens. Therefore, they have evolved an innate immune system based upon complicated and efficient defense mechanisms, either constitutive or inducible. Plant defense responses are triggered by elicitors such as microbe-associated molecular patterns (MAMPs). These components are recognized by pattern recognition receptors (PRRs) which include plant cell surface receptors. Upon recognition, PRRs trigger pattern-triggered immunity (PTI). Ethylene Inducing Xylanase (EIX) is a fungal MAMP protein from the plant-growth-promoting fungi (PGPF)–Trichoderma. It elicits plant defense responses in tobacco (Nicotiana tabacum) and tomato (Solanum lycopersicum), making it an excellent tool in the studies of plant immunity. Xylanases such as EIX are hydrolytic enzymes that act on xylan in hemicellulose. There are two types of xylanases: the endo-1, 4-β-xylanases that hydrolyze within the xylan structure, and the β-d-xylosidases that hydrolyze the ends of the xylan chain. Xylanases are mainly synthesized by fungi and bacteria. Filamentous fungi produce xylanases in high amounts and secrete them in liquid cultures, making them an ideal system for xylanase purification. Here, we describe a method for cost- and yield-effective xylanase production from Trichoderma using wheat bran as a growth substrate. Xylanase produced by this method possessed xylanase activity and immunogenic activity, effectively inducing a hypersensitive response, ethylene biosynthesis, and ROS burst.


2021 ◽  
Vol 22 (10) ◽  
pp. 5064
Author(s):  
Qinghua Chen ◽  
Linghui Guo ◽  
Yanwen Yuan ◽  
Shuangling Hu ◽  
Fei Guo ◽  
...  

Histone methylation plays an important regulatory role in the drought response of many plants, but its regulatory mechanism in the drought response of the tea plant remains poorly understood. Here, drought stress was shown to induce lower relative water content and significantly downregulate the methylations of histone H3K4 in the tea plant. Based on our previous analysis of the SET Domain Group (SDG) gene family, the full-length coding sequence (CDS) of CsSDG36 was cloned from the tea cultivar ‘Fuding Dabaicha’. Bioinformatics analysis showed that the open reading frame (ORF) of the CsSDG36 gene was 3138 bp, encoding 1045 amino acids and containing the conserved structural domains of PWWP, PHD, SET and PostSET. The CsSDG36 protein showed a close relationship to AtATX4 of the TRX subfamily, with a molecular weight of 118,249.89 Da, and a theoretical isoelectric point of 8.87, belonging to a hydrophilic protein without a transmembrane domain, probably located on the nucleus. The expression of CsSDG36 was not detected in the wild type, while it was clearly detected in the over-expression lines of Arabidopsis. Compared with the wild type, the over-expression lines exhibited lower hyperosmotic resistance by accelerating plant water loss, increasing reactive oxygen species (ROS) pressure, and increasing leaf stomatal density. RNA-seq analysis suggested that the CsSDG36 overexpression caused the differential expression of genes related to chromatin assembly, microtubule assembly, and leaf stomatal development pathways. qRT-PCR analysis revealed the significant down-regulation of stomatal development-related genes (BASL, SBT1.2(SDD1), EPF2, TCX3, CHAL, TMM, SPCH, ERL1, and EPFL9) in the overexpression lines. This study provides a novel sight on the function of histone methyltransferase CsSDG36 under drought stress.


2004 ◽  
Vol 85 (1) ◽  
pp. 33-37 ◽  
Author(s):  
Magali Merkx-Jacques ◽  
Jacqueline C. Bede

Abstract Plants exhibit remarkable plasticity in their ability to differentiate between herbivorous insect species and subtly adjust their defense responses to target distinct pests. One key mechanism used by plants to recognize herbivorous caterpillars is elicitors present in their oral secretions; however, these elicitors not only cause the induction of plant defenses but recent evidence suggests that they may also suppress plant responses. The absence of “expected changes” in induced defense responses of insect-infested plants has been attributed to hydrogen peroxide produced by caterpillar salivary glucose oxidase (GOX). Activity of this enzyme is variable among caterpillar species; it was detected in two generalist caterpillars, the beet armyworm (Spodoptera exigua) and the bertha armyworm (Mamestra configurata), but not in other generalist or specialist caterpillar species tested. In the beet armyworm, GOX activity fluctuated over larval development with high activity associated with the salivary glands of fourth instars. Larval salivary GOX activity of the beet armyworm and the bertha armyworm was observed to be significantly higher in caterpillars reared on artificial diet as compared with those reared on Medicago truncatula plants. This implies that a factor in the diet is involved in the regulation of caterpillar salivary enzyme activity. Therefore, plant diet may be regulating caterpillar oral elicitors that are involved in the regulation of plant defense responses: our goal is to understand these two processes.


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