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題名 | The Structural, Biochemical, and Genetic Characterization of a New Radiation-induced, Variegated Leaf Mutant of Soybean [Glycine max (L.) Merr.]=大豆葉斑紋突變E25-10葉片構造遺傳與生化之研究 |
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作者姓名(中文) | 程臺生; | 書刊名 | Proceedings of the National Science Council : Part B, Life Science |
卷期 | 23:1 1999.01[民88.01] |
頁次 | 頁27-37 |
分類號 | 373.4 |
關鍵詞 | 大豆葉斑紋; 突變; 葉片構造; 遺傳; 生化; Photosystem Ⅰ; Pigment-protein complexes; Soybean; Variegated leaf mutant; |
語文 | 英文(English) |
中文摘要 | 本研究採用生理、生化與遺傳方法對經放射線鈷六十誘發之大豆葉斑紋突變種E25 -10 進行分析。 E25-10 葉片的黃色斑紋葉肉細胞內含缺失的葉綠體,由葉表顏色深淺不一 且挾雜相間, 及其缺失葉綠體所位在葉肉細胞層的排列方式判斷,E25-10 可能具有自發回 復突變的能力。在電子顯微鏡下觀察缺失的葉綠體,其葉綠餅膜以相互平行、無或甚少重疊 方式排列,不含澱粉粒,而小囊袋或油狀體則可見。遺傳研究則顯示 E25-10 為核基因突變 體。 E25-10 黃色斑紋葉肉組織所含葉綠素較正常葉肉組織減少 70-80 %, 雖然其葉綠素 a/b 比值仍維持不變, 但是光譜分析顯示在波長 400-500nm 間,其葉綠素吸收光譜與正常 葉肉組織所含葉綠素並不相同。這種葉綠素含量減少與吸收光譜改變,應與缺失的葉綠體內 缺損某些光合作用複合體有關。綠色膠體電泳分析顯示,在缺失的葉綠體內四個主要的色素 -蛋白複合體( CP1, LHCP ��,LHCP �援M CPa )皆不存在。 LDS-PAGE 電泳分析發現,至 少有 5-6 個葉綠餅膜蛋白在缺失的葉綠體內不可見, 其分子量分別是 51,44,25,15,13 及 12kDa。在黃色斑紋葉肉細胞內,葉綠體或細胞核基因表現研究結果顯示,葉綠體基因 psaA ( PSI ),rbcL,16S rRNA 與細胞核基因 rbcS,cab 都有不同程度的衰減。 綜合以上結 果,雖然 E25-10 缺失的葉綠體內甚少重疊之葉綠餅膜,但是我們認為突變發生的部位仍以 光反應系統I的可能性較大。 |
英文摘要 | A variegated leaf mutant in soybean [Glycine max (L.) Merr.] has been identified and characterized. E25-10 was derived by exposure of seeds of the "Williams'82" cultivar to a-radiation. In this mutant, yellow leaf sectors contain defective chloroplasts, in which the thylakoid membranes are presented as long, parallel structures with little or no overlap. No starch grains have been detected in the mutant chloroplasts. Small vesicles and plastoglobuli can be found within the defective chloroplasts. Genetic studies revealed that a single nuclear-encoded gene is responsible for the mutation in E25-10. The total chlorophyll content is reduced in yellow leaf tissue by 70-80%. However, the chlorophyll a/b ratio is not altered. The absorbance spectrum of pigments in the mutant leaf tissue differed from that of the green extracts in the range of 400-500 nm. This reduction in total chlorophyll and the change in the absorbance spectrum pattern in the yellow tissue is related to a loss of certain photosynthetic complexes. Green gel analysis revealed that four major pigment-protein complexes (CP1, LHCP1, LHCP2, and CPa) of the thylakoid membranes were absent in the E25-10 mutant. Lithium dodecyl sulphate polyacrylamide gel analysis showed that at least 5-6 polypeptides (51, 44, 25, 15, 13, and 12 kDa) were missing in the thylakoid membranes of chloroplasts from the yellow tissue. Changes in chloroplast- and nuclear-encoded gene message levels were detected. The psaA transcripts which code for the P700 apoprotein in PSI were reduced in chloroplasts from the E25-10 mutant yellow tissue. The levels of the large subunit of ribulose bisphosphate carboxylase (rbcL) and light harvest complex protein (LHCP) of PSII mRNA appeared to be reduced slightly in the mutant plants. However, a much more significant reduction in the 16S rRNA and the small subunit of ribulose bisphosphate carboxylase (rbcS) expression was detected in the yellow leaf sectors. Our results suggest that the possible lesion in E25-10 is located in the photosystem I even though fewer grana were observed in the defective chloroplasts. |
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