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Effects of Exogenous Nitric Oxide Donor Sodium Nitroprusside on β-Amylase during Early Stage of Seed Germination in Triticum Aestivum L.

Author: SunYongGang
Tutor: ShenWenZuo
School: Nanjing Agricultural College
Course: Biochemistry and Molecular Biology
Keywords: Germination beta - amylase NO Subcellular localization Wheat seeds
CLC: S512.1
Type: Master's thesis
Year: 2009
Downloads: 52
Quote: 0
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Abstract


Starchy crops wheat (Triticum aestivum L.) study found that beta-amylase-specific inhibitor of alpha-cyclodextrin wheat seed soaking treatment, found to significantly inhibit the germination of wheat seeds, description beta- amylase in wheat seed germination and early plays an important role (at least the first 24 hours); 0.5 mmol L-1 SNP treatment alone can significantly improve the germination of wheat seeds and two common wheat seeds found that alpha-cyclodextrin SNP treatment also have some inhibitory effect. Further analysis of amylase activity and starch content of wheat seed SNP treatment can significantly improve the germination of the early wheat seed total amylase activity (germination six hours most significant difference), and seed starch content compared with the control treatment of low. Not measured during this process the alpha-amylase activity, substrate specificity and the use of beta-amylase activity was measured, the results showed that SNP treatment can improve the wheat seeds vivo total beta-amylase activity. To having reducing the SNP may be a non-active conjugated beta-amylase converted to active free beta-amylase, and thus freedom and type-binding beta-amylase were measured and found SNP processed mainly by improving The free-type beta-amylase activity and to improve the total beta-amylase activity; contrast, the reducing agent beta-mercaptoethanol treatment is simply to freedom transformation through a combination of type and increase the total beta-amylase activity . Combination the NO scavenger cPTIO jointly deal, cPTIO reversed to some extent, the SNP on the activity of beta-starch increased, while no significant effect on reducing agent beta-mercaptoethanol treatment. Further found by semi-quantitative RT-PCR test the SNP can improve beta-amylase gene transcripts by Western blotting SNP treatment induced the expression of beta-amylase protein, the SNP and the NO-specific scavenger cPTIO and transcription inhibitor actinomycin D, treatment can significantly reverse the above changes in transcript and protein levels. Immune electron microscopy subcellular localization results show that beta-amylase protein was mainly located in the wheat germ protein storage, and may be attached to the starch granules, SNP treatment increased the protein content and distribution of beta-amylase, at the same time reducing agent beta - mercaptoethanol processing is also supported by a similar result, but less than the SNP the specific SNP NO scavenger cPTIO of the processing result is consistent with the results of the above tests, wherein the beta-amylase protein content and distribution restored to the level of the control . The above results show that exogenous NO donor SNP early induced de novo synthesis of beta-amylase protein to improve the free beta-amylase activity in wheat seed germination.

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CLC: > Agricultural Sciences > Crop > Cereal crops > Wheat > Wheat
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