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Phytochemicals and antioxidative enzymes defence mechanism on occurrence of yellow vein mosaic disease of pumpkin (Cucurbita moschata).

ABSTRACT: Pumpkin (Cucurbita moschata) samples showing yellow vein mosaic disease in Varanasi region were identified with begomovirus infection using PCR amplification. A sequencing analysis of the full genome revealed that it is a strain of Tomato leaf curl Palampur virus (GenBank ID. FJ931537). Phytochemical composition and antioxidative enzyme levels were compared in infected and healthy plants. The study revealed that the amount of total protein declined in the infected leaves but elevated up to 135 % in the fruits of infected plants, whereas vitamin C and antioxidants declined in infected leaves as well as fruits. There was substantial increase in total phenol content in leaves (72 %) and fruits (300 %) of infected plants. In infected samples, substantial increase in activities of superoxide dismutases (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and catalase (CAT) was observed as compared to the uninfected control plants. The native PAGE showed alterations in the intensities of isozyme bands in the infected plants. The APX, GPX, CAT, SOD and glutamate dehydrogenase (GDH) bands were intense in the infected plants, whereas the GR isozyme showed reduced intensity in diseased plants.

SUBMITTER: Jaiswal N

PROVIDER: S-EPMC3723865 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Background:Quantitative real-time PCR (qRT-PCR) is a commonly used high-throughput technique to measure mRNA transcript levels. The accuracy of this evaluation of gene expression depends on the use of optimal reference genes. Cucumber-pumpkin grafted plants, made by grafting a cucumber scion onto pumpkin rootstock, are superior to either parent plant, as grafting conveys many advantages. However, although many reliable reference genes have been identified in both cucumber and pumpkin, none have been obtained for cucumber-pumpkin grafted plants. Methods:In this work, 12 candidate reference genes, including eight traditional genes and four novel genes identified from our transcriptome data, were selected to assess their expression stability. Their expression levels in 25 samples, including three cucumber and three pumpkin samples from different organs, and 19 cucumber-pumpkin grafted samples from different organs, conditions, and varieties, were analyzed by qRT-PCR, and the stability of their expression was assessed by the comparative ?Ct method, geNorm, NormFinder, BestKeeper, and RefFinder. Results:The results showed that the most suitable reference gene varied dependent on the organs, conditions, and varieties. CACS and 40SRPS8 were the most stable reference genes for all samples in our research. TIP41 and CACS showed the most stable expression in different cucumber organs, TIP41 and PP2A were the optimal reference genes in pumpkin organs, and CACS and 40SRPS8 were the most stable genes in all grafted cucumber samples. However, the optimal reference gene varied under different conditions. CACS and 40SRPS8 were the best combination of genes in different organs of cucumber-pumpkin grafted plants, TUA and RPL36Aa were the most stable in the graft union under cold stress, LEA26 and ARF showed the most stable expression in the graft union during the healing process, and TIP41 and PP2A were the most stable across different varieties of cucumber-pumpkin grafted plants. The use of LEA26, ARF and LEA26+ARF as reference genes were further verified by analyzing the expression levels of csaCYCD3;1, csaRUL, cmoRUL, and cmoPIN in the graft union at different time points after grafting. Discussion:This work is the first report of appropriate reference genes in grafted cucumber plants and provides useful information for the study of gene expression and molecular mechanisms in cucumber-pumpkin grafted plants.

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Phytochemicals and antioxidative enzymes defence mechanism on occurrence of yellow vein mosaic disease of pumpkin (Cucurbita moschata).

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