熱休克蛋白基因與過氧化氫酵素基因轉移到甘藍葉綠體之研究

Studies on Transformation of Heat Shock Protein Gene and Catalase Gene into the Chloroplast of Cabbage (Brassica oleracea L. var. capitata L.)

陳志宏(Z. H. Chen) ； 劉程煒(C. W. Liu) ； 尤進欽(J. C. Yiu) ； 陳良築(L. J. Chen) ； 曾夢蛟(M. J. Tseng)

《植物種苗》 8卷1期 (2006/03) Pp. 23-45

https://doi.org/10.30077/SN.200603.0003

摘要

本研究分別將以prrn為啟動子的番茄hsc70與結球白菜cat78基因構築至蘑菇蕓苔屬葉綠體基因轉殖之通用轉殖載體(pASCC201)，成為pASCCHSC及pASCCCAT之蕓苔屬葉綠體基因轉殖載體。再以基因槍法將其分別轉移到甘藍葉片、子葉及下胚軸，經誘導再生植株。以基因槍法轉殖之培殖體，以低濃度的抗生素(spectinomycin)持續篩選4週復，可獲得轉殖的再生植株。以PCR及南方墨點分析hsc70、cat78之再生甘藍植株之葉片結果約可偵測到轉殖之aadA、hsc70與cat78基因的存在，顯示hsc70、cat78基因已轉移到甘藍之葉綠體基因組。以北方墨點分析轉殖甘藍植株可偵測到轉移之hsc70 mRNA的表現，以西方墨點分析Hsc70之結果顯示在轉殖甘藍有較強的Hsc70表現。轉殖甘藍植株經由42℃高溫處理三天後，較未轉殖對照組呈現極耐高溫之效果。

關鍵字

葉綠體基因轉移 ； 熱休克蛋白基因 ； 過氧化氫酵素基因 ； 甘藍

並列摘要

The objective of this study was to explore the possibility for improvement of cabbage with heat and oxidative resistance by chloroplast gene transformation. The heat shock cognate protein gene (hsc70) of tomato and catalase 78 gene (cat78) of Chinese cabbage driven by prrn promoter were constructed into the plastid-specific vector of Brassica, pASCC201; to form the pASCCHSC and pASCCCAT, respectively. The hsc70, and cat78 genes were transferred into the chloroplast of cabbage via particle gun mediated transformation. Regenerated plantlets were screened by resistance to spectinomycin antibiotics. The results of PCR and Southern hybridization analysis indicated that the aadA, hsc70, and cat78 genes were integrated into the chloroplast genome of cabbage. Furthermore, the expression of hsc70 mRNA and Hsc70 protein were detected in the hsc70 gene transformed cabbages by northern blot analysis and western blot analysis, respectively. After 3 days of 42℃ treatments, higher degree of heat resistance was found in hsc70 gene transformed cabbage than those of non-transformed plants.

並列關鍵字

chloroplast transformation ； heat shock protein gene ； catalase gene ； cabbage