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研究生:張佳欣
研究生(外文):Chang, Chia-Hsin
論文名稱:高粱可溶性澱粉合成�A活性及其分子特性
論文名稱(外文):Activity and molecular characterization of sorghum soluble starch synthase
指導教授:謝兆樞謝兆樞引用關係
指導教授(外文):Hsieh Jaw-Shu
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:農藝學系
學門:農業科學學門
學類:一般農業學類
論文種類:學術論文
論文出版年:1997
畢業學年度:85
語文別:中文
論文頁數:84
中文關鍵詞:高粱可溶性澱粉合成
外文關鍵詞:sorghumsoluble starch synthase
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本論文主要在探討高粱未成熟種子胚乳及成熟葉片中,可溶性澱粉合成
活性在高溫下所受的影響。實驗結果證實,不論在高粱未成熟種子胚乳或
成熟之完全展開葉中的可溶性澱粉合成,其活性在溫度升至35℃後,均
有降低的現象,特別在蠟質品系III wx中,酵素活性降低得相當迅速。由
胚乳的試驗組及葉片的試驗組的比較中可發現,葉片中的可溶性澱粉合成
對溫度的敏感性較胚乳中者為高。另外,由EMS誘導突變的高粱蠟質品
系EMS-wx,其葉片中的可溶性澱粉合成活性在溫度升至30℃,活性即迅
速的下降,這在熱帶禾穀類作物中,是一個較不常見的現象。為了進行高
粱可溶性澱粉合成cDNA殖系選殖,藉由比對水稻可溶性澱粉合成與高
粱Wx蛋白序列中較為保守的區域設計四支引子,經由PCR合成高粱可溶性
澱粉合成的同源性探針,再以此探針作為篩選高粱可溶性澱粉合成殖
系之用。且該PCR增殖產物業已經由Southern blot雜合反應,確認其不同
於高粱Wx cDNA片段。但由於篩選時未注意溶菌斑形成力價不足,以致篩
選效率低落,而沒有選到正確殖系。在SDS-PAGE及西方墨點轉印的分析上
,我們發現不論由非蠟質及蠟質品系未成熟種子中抽取的可溶性蛋白,皆
可被高粱蠟質蛋白之抗血清辨認出一分子量約為70kD的蛋白分子,須進一
步的實驗確定,該蛋白是否為高粱胚乳中可溶性澱粉合成。
In this thesis, we investigated the effect of high temperature
on the activity of soluble starch synthase in sorghum immature
seed endosperm and leaf. The results showed that the enzyme
activity of the soluble starch synthase, in terms of 14C-glucose
incorporation rate, in both organs would be decreased under high
temperature treatment, the decrement was especially significant
in waxy sorghum lines, III wx. Our result also showed that, the
enzyme in leaves was more sensitive to the alternation of
temperature than that in the endosperm. The activity of soluble
starch synthase in leaves of the EMS induced waxy mutant EMS-wx
line went quickly down under temperature over 30℃. This was not
a common phenomenon in tropical crops.In order to clone the cDNA
encoding sorghum soluble starch synthase, we used four primers,
which located on the conserved regions of the open reading frame
of rice soluble starch synthase and sorghum Wx cDNA sequence, to
synthesize homologous probes by PCR. The amplified products were
not the Wx-related product, as confirmed by Southern blot
analysis, and they were used as the probes for cDNA library
screening. The plaques forming unit was not high enough to make
the screening efficient. Suitable condition still need to be
tested. By SDS-PAGE and western blot analysis, we identified a
70kD, recognized by anti-Wx protein antiserum, that was present
both in the non-waxy and waxy sorghum immature endosperm.
Whether the protein can be clarified as a isozyme of sorghum
soluble starch synthase or not need more experimental data to
confirm with.
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