Parinthawong, Nonglak

Preferred name

Alternative Name

Parinthawong, N.

Main Affiliation

School of Agricultural Technology

nonglak.pa@kmitl.ac.th

Now showing 1 - 10 of 27

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Isolation and expression analysis of the Flavanone 3-Hydroxylase genes in Lotus (Nelumbo nucifera gaertn.), Waterlily (Nymphaea sp.) and transient silencing in Waterlily
(2017-04-01)
Chaipanya, Chaivarakun
;
Saetiew, Kanjana
;
Arunyanart, Sumay
;
Parinthawong, Nonglak
Flavanone 3-hydroxylase (F3H) converts flavanones from dihydroflavonols, which leads to production of flavonoid compounds via the anthocyanin biosynthesis pathway in plants. In this study, the F3H gene was isolated from lotus (Nelumbo nucifera Gaertn.) cv. Buntharik (white petal lotus), cv. Satabankacha (pink petal lotus), and waterlily (Nymphaea sp.) var. St. Louis Gold by reverse transcription PCR (RT-PCR). The open reading frames (ORF) of three cultivars’ genes were 1,134 bp in length, encoding a predicted protein of 377 amino acids. Their nucleotide sequences were identical, and the amino acid sequence shared high homology to F3H from different plant species. Expression of F3H was specifically regulated in petals and stamens, while less expression was found in leaf tissue of waterlily variety St. Louis Gold. The correlation of F3H expression according to specific colouration was performed in waterlily. The F3H gene was more highly expressed in decreasing order of red, purplish blue, and yellow petals when compared using semi quantitative PCR (sqPCR). Gene regulation according to flowering stage and pigmentation was determined in lotus. The F3H expression was slightly diminished in petals of cultivar Satabankacha at the fully-opening stage, whereas it was detected in the cultivar Buntharik only when white petals were tinted with pink. An RNAi gene-silencing vector, pJA8F3H, encoding a hairpin F3H RNA, was introduced to waterlily petals using the Agrobacterium infiltration method, and the F3H expression was analysed at 1 and 3 days post infiltration (dpi) by sqPCR. The results showed that the F3H expression was down-regulated at 3 dpi in flowers tested of the red petal variety and purplish blue petal variety compared to controls. The results confirmed that pJA8F3H is efficient and could be used as a transformation vector to transiently suppress F3H expression in waterlily or lotus.
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Mating type and genetic diversity analysis of Pyricularia oryzae collected from Thai rice varieties
(2018-12-01)
Tansian, P.
;
Parinthawong, N.
Rice blast disease caused by Pyricularia oryzae, is one of the most damaged of rice production worldwide. The fungi are a great diversity on both genotype and pathotype. The genetic relationship of P. oryzae collected in Thailand during year 2016 and 2017 was analyzed. Rice plants appeared blast symptom were collected from epidemic areas and was isolated by single spore. All isolates were identified for mating type using MAT1-1 and MAT1-2 primer sets. In this result, the mating type MAT1-2 was mostly found in Thailand and only about 7.7 % of MAT1-1 was found in the population. In this observation, both MAT1-1 and MAT1-2 were found from isolates collected from the same location in Phetchabun province. Therefore, the genetic analysis of 14 Magnaporthe grisea using microsatellite (MGM) markers was cluster-analyzed using UPGMA method of the SHAN program. The result showed cluster analysis was separated the population into 11 groups. Group 1 to 7 showed the percentage of the population with 3.8, 5.8, 36.5, 13.5, 3.8, 25.1 and 3.8, respectively. For group 8-11, the rice blast pathogen from North and South of Thailand were separated into single isolate group. Group 6 was the highest diversity of province numbers which obtained from diverse rice varieties that showed similar genotype but difference in pathogenicity. The difference isolates of fungi could infect the same rice variety. Genetic diversity and determination of mating type of rice blast populations in Thailand are needed to study and useful for predicting an epidemic of rice blast disease and selecting the appropriate isolates for blast breeding programs.
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Transformation of antisense dihydroflavonal 4-reductase (DFR) into sacred lotus 'Buntharik' using Agrobacterium-mediated gene transfer
(2014-03-13)
Saetiew, K.
;
Leethaweesup, W.
;
Parinthawong, N.
;
Arunyanart, S.
Transformation of antisense dihydroflavonal 4-reductase (DFR) into 'Buntharik' sacred lotus (Nelumbo nucifera Gaertn.) using Agrobacterium-mediated gene transfer was experimented. The A. tumefaciens strain EHA105 harbored two binary vectors. The pCAMBIA2301anti-DFR plasmid contained neomycin phosphotransferase (NPTII) as a selectable marker gene, the antisense DFR as an inserted gene and the β-glucuronidas gene (GUS) as a reportor gene. The pBI121anti-DFR plasmid contained NPTII gene and the antisense DFR. Twomonths-old calli regenerated from apical buds were used as plant materials. The plant materials were soaked in the Agrobacterium suspension for 10 or 30 min. The calli were co-cultivated for two days in the darkness. Treated calli were selected on solid MS medium containing 50 mg L<sup>-1</sup> kanamycin and 250 mg L<sup>-1</sup> cefotaxime. The calli were transferred to the same medium every two weeks for eight weeks. The survived calli regenerated shoots on MS medium containing a combination of 0.54 μM NAA and 4.44 μM BA. Both plasmids gave the best callus growth, maximum callus size and the survival percentage when soaked for 10 min. GUS bioassay was used to verify the presence of GUS gene in petioles and leaves of transgenic plants. It was found that petioles and leaves of transformants which were transformed with pCAMBIA2301anti-DFR by soaking for 30 min showed the highest percentage of blue spots on explants. Of all 14 PCR positive clones, 5 clones showed GUS gene, 11 clones contained NPTII gene and 6 clones had DFR gene.
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Transformation RNA interference-mediated silencing of the Flavanone 3-Hydroxylase gene in Lotus (Nelumbo nucifera Geartn. cv. Buntharik)
(2020-07-01)
Saetiew, K.
;
Konee, C.
;
Parinthawong, N.
;
Arunyanart, S.
RNA interference mediated-silencing of the F3H gene in the lotus (Nelumbo nucifera Geartn.) cv. Buntharik inhibited the anthocyanin pathway after gene transformation. The plasmid pJA8F3H contained the inverted repeats of F3H fragments, bar gene as a selectable marker in plants, which is resistant to basta (glufosinate amonium). The transformation of particle bombardment was obtained with multiple shoots of lotus. F3H gene expression analysis showed that expression of F3H decreased in transgenic lotus when compared with the control. On the first and the second test, 5 transgenic lotus of the relative expression F3H gene was detected with 0.0496 and 0.466 times, respectively. The inhibition of gene expression decreased when they were cultured for a long time. The expression F3H gene was detected by RT-PCR, and real-time PCR resulted in consistent decreasing in the transgenic lotus compared with the non-transgenic lotus, which was able to inhibit the expression F3H gene.
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Non-Pathogenic Pythium Induced the Resistance in Hydroponic Lettuce Against Root Rot Disease
(2024-05-01)
Talubnak, Chulalak
;
Parinthawong, Nonglak
;
Schoonbeek, Henk Jan
;
Jaenaksorn, Tanimnun
Hydroponics is a common soilless type of culture. It is clean and gives high-yield crop production. Lettuce is the most widely grown leafy vegetable in hydroponics. The Pythium species causes root rot, a serious disease in lettuce. In this study, we investigated an effective elicitor to promote resistance in lettuce after pathogen infection. To evaluate the plant defense gene response profile, the expression stability of five candidate genes was studied using RT-qPCR. The LsPR1 and LsLTC2 gene expression were higher in lettuce leaves treated with non-pathogenic Pythium PyASR23 than in roots compared to untreated plants. To assess the disease, the rate of relative gene expression of LsLTC2, LsPR-1b like, LsERF, LsAOS, and LsDEF was associated with Pythium root rot pathogenesis. As a result, it was revealed that alterations in these gene expressions were a defense response to Pythium infection, which increased after pre-treatment with various elicitors for 24 hours. When the chemically stimulated lettuce was infected with pathogenic PySR31, the expression of these genes increased dramatically. These results suggested that non-pathogenic PyASR23 might be employed as an elicitor in lettuce roots and contribute to disease control by activating plant defense gene expression against the Pythium pathogen.
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Genetic mapping of leaf blast resistance gene in landrace rice cultivar ‘GS19769’
(2015-08-31)
Parinthawong, Nonglak
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Tansian, Pennapar
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Sreewongchai, Tanee
The rice blast fungus, Magnaporthe oryzae, is highly varied and therefore overcomes resistance in a few years. Rice cultivars with the ability to resist different blast races are therefore required. In this study, a resistance gene of landrace rice cultivar ‘GS19769’ is identified using a mix of 19 blast isolates collected from several epidemic areas in Thailand. The selected cultivar was fertilised with the blast susceptible variety, Khao Dok Mali 105 (KDML105), to generate the mapping population. Segregation analysis in the F<inf>2</inf> population shows that ‘GS19769’ contains more than 1 resistance gene, as the Chi-square test for segregation of resistance and susceptibility does not fit the ratio of 3:1. The bulk segregant analysis by simple sequence repeat (SSR) markers shows that the identified resistance gene is linked to the SSR markers RM224 and RM144 on chromosome 11. Analysis of 15 F<inf>2</inf> plants susceptible to the blast was conducted. The gene was mapped to RM224 and RM144 at the same distance of 20 cM.
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High nucleotide sequence variation of avirulent gene, AVR-Pita1, in Thai rice blast fungus population
(2020-12-01)
Damchuay, Katanyutita
;
Longya, Apinya
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Sriwongchai, Tanee
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Songkumarn, Pattavipha
;
Parinthawong, Nonglak
Rice blast disease, caused by Magnaporthe oryzae, is one of the most importance diseases of rice production worldwide. The key role of defense mechanism to combat this fungus in rice follows the gene-for-gene concept, which a plant resistant (R) gene product recognizes a fungal avirulent (AVR) effector and triggers the hypersensitive response. However, the AVR genes have been shown to be rapidly evolving resulting in high level of genetic diversity. The aims of this study were to examine the nucleotide sequence variation of AVR-Pita1 gene in Thai rice blast isolates and to identify the severity of blast disease using isogenic line of Pita gene. Seventy-six rice blast isolates collected from different parts of Thailand were used. Gene specific primers for AVR-Pita1 gene coding sequence were designed and used for identifying the genetic diversity of AVR-Pita1 gene by PCR amplification and sequencing. The obtained sequences were analysed for genetic variation and genetic relationship. Our results revealed the association between the sequence variations of AVR-Pita1 and selective forces from Pita gene. This phenomenon demonstrated the coevolution between rice blast resistant gene in rice and avirulent gene in blast fungus. The information about variation and evolutionary mechanisms of AVR gene obtained from this study can be used in rice blast resistant breeding programme.
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Identification of blast-resistant varieties from Landrace, improved and wild species of rice
(2013-07-18)
Salih, Abdu
;
Sreewongchai, Tanee
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Sripichitt, Prapa
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Parinthawong, Nonglak
An experiment was conducted to identify rice varieties resistant to blast disease. Samples of 311 genetically diversified varieties/accessions comprised of landrace (263), improved (43) and wild (5) rice varieties/accessions were provided by the National Rice Gene Bank of Thailand. The screening for blast at the seedling stage was done using 29 diversified blast isolates. The blast isolates were collected from seven provinces (Phitsanulok, Ubon Ratchathani, Khon Kean, Chiang Rai, Nong Khai, Chaiyaphum and Udon Thani) of Thailand. The results indicated a total of 35 varieties/accessions (25 landrace, 9 improved and 1 wild) were resistant to all tested blast isolates. The 25 resistant landrace varieties were collections from Northern (10), Northeastern (9) and Southern (6) Thailand. Moreover four of the resistant landrace varieties (GS23107, GS19769, GS20874 and GS23774) were highly resistant with no symptom of the disease. From this study, it can be suggested that the resistant landraces from the Southern, Northern and Northeastern regions could be used as sources of resistant varieties in designing future breeding programs aimed at developing disease-resistant genotypes.
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Protein hydrolysates from agricultural wastes for plant bacterial disease control
(2022-03-01)
Ditsawanon, T.
;
Parinthawong, N.
;
Phaonakrob, N.
;
Roytrakul, S.
Agricultural wastes, agro-industrial wastes and fishery wastes were collected and the protein hydrolysates were obtained with pepsin. Antibacterial activity of smaller than 3 kDa protein hydrolysates was determined against the plant pathogenic bacteria; Xanthomonas citri, Ralstonia solanacearum, Burkholderia cepacia and also against plant growth promoting rhizobacteria (PGPRs); Bacillus subtilis, Pseudomonas aeruginosa and P. fluorescens. Coconut residues (agro-industrial waste from coconut milk production), peanut seed coat (from peanut-based snack production) and rice straw (waste from rice farms) showed antimicrobial activity against X. citri, R. solanacearum and B. cepacia with higher than 74% inhibition. Coconut residue also increased growth of PGPRs, B. subtilis and P. fluorescens. Further protein hydrolysates from Nile tilapia (Oreochromis niloticus) and snake-head fish (Clarias batrachus) fin increased growth of all PGPRs.
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Genetic variation of avirulence genes (AVR-Pi9, AVR-Pik, AVR-Pita1) and genetic diversity of rice blast fungus, Pyricularia oryzae, in Thailand
(2022-02-01)
Sutthiphai, Thanathip
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Damchuay, Katanyutita
;
Neupane, Ram Chandra
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Longya, Apinya
;
Sriwongchai, Tanee
Genetic variation of the rice blast (Pyricularia oryzae) population in Thailand was investigated based on the nucleotide sequence of three avirulence genes, AVR-Pi9, AVR-Pik, and AVR-Pita1. Sixty rice blast isolates were collected from rice-growing areas around Thailand. Gene-specific primers were used to amplify these AVR genes and AVR-Pi9, AVR-Pik, and AVR-Pita1 were detected in 60, 57, and 23 isolates, respectively. Based on the AVR-Pi9 sequences, we identified one rice blast isolate containing an amino acid change from glutamic acid to aspartic acid. Moreover, two rice blast isolates had identical sequences to the rice blast strain originating in Japan, indicating a potential movement of this isolate from Japan to Thailand. Three AVR-Pik alleles were found, including AVR-PikA (3.51%), AVR-PikD (71.93%), and isolates with two copies of AVR-PikD and AVR-PikF (24.56%). AVR-PikA and AVR-PikF are virulent to Thai rice variety Jao Hom Nin. Six haplotypes of AVR-Pita1 were identified with one deletion and 12 amino acid substitutions. This study revealed that different AVR genes in Thai rice blast populations have different levels of genetic variation: AVR-Pi9 and AVR-Pik genes have a relatively low genetic diversity, while the AVR-Pita1 gene has high genetic diversity. We found AVR-Pi9 was not under selection pressure, while AVR-Pita1 was under purifying selection pressure. In addition, geographic location has influenced the distribution of genetic variation of AVR-Pita1. The information obtained from this study is valuable for the future development of breeding strategies for rice blast resistance in Thailand.

Parinthawong, Nonglak

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