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    醫用卡那霉素在轉基因抗蟲棉鑒定中的應用

    原文作者:馮振秀、朱波、曹陽

    摘要 利用醫用硫酸卡那霉素對不同品種的棉花葉片進行處理,研究不同濃度醫用硫酸卡那霉素溶液、不同棉花品種葉片對卡那霉素處理的外觀變化。結果表明:不同棉花品種葉片對卡那霉素處理呈相同的變化趨勢。當卡那霉素濃度達到5 000 mg/l時,檢測準確度最好。
       關鍵詞 轉基因;棉花;醫用卡那霉素;濃度 [論文網]
      中圖分類號 s562 文獻標識碼 a 文章編號 1007-5739(2013)20-0023-01
      由于轉基因抗蟲棉花品種在農業生產上大面積應用和各地引種工作的日益頻繁,在人們受益于轉基因抗蟲棉這項高新技術的同時,也存在著已知的或潛在的生態安全風險。因此,必須采取有效措施控制和預防這種風險。
       利用實驗室技術進行轉基因抗蟲棉花品種檢測較繁瑣,操作不便[1-3]。而田間用醫用卡那霉素直接對棉花葉片進行抗性鑒定是一項方便、有效的技術[4]。但農作物對卡那霉素的敏感度存在差異,而棉花葉片對卡那霉素存在抗性反應。因此,研究棉花葉片對卡那霉素處理的反應,進而發展、推廣應用該技術,以提高轉基因抗蟲棉檢測、鑒定的準確性。
       1 材料與方法
      1.1 試驗材料
      參試棉花品種:非轉基因品種新陸早36號、自育品系y153-2和35-2;轉基因抗蟲對照品種為抗蟲5號、抗蟲10號。硫酸卡那霉素為徐州萊恩藥業有限公司生產的2 ml:0.5 g(50萬u)醫用注射液。www.zhidachem.com一次性醫用注射器、醫用脫脂棉、蒸餾水(或純凈水)。
       1.2 試驗方法
      試驗前將醫用卡那霉素注射液用蒸餾水分別配制成1 000、2 000、3 000、4 000、5 000 mg/l 5個不同濃度的處理。試驗時,用醫用注射器吸取不同濃度的卡那霉素溶液,拔除針頭,塞上醫用脫脂棉,在棉花生長的四葉期,涂抹棉花新展開倒1葉上的相同部位,直徑1~2 cm,以濕潤為準。每個參試品種,利用不同濃度的卡那霉素溶液各處理60株,并在田間做好相應的標記。5 d后觀察各處理涂抹部位的葉色反應,并做好相應數據的記載和后期處理、分析工作。應選擇在清晨或傍晚涂抹,避開高溫、蒸發劇烈和大風或降雨等異常天氣。
       2 結果與分析
      2.1 棉花葉片卡那霉素處理后外觀表現的篩選標準
      根據田間觀察棉花葉片對卡那霉素處理后的外觀表現:將卡那霉素溶液涂抹部位的葉色與周邊正常葉色存在明顯差異,葉色由綠轉黃甚至出現破損的情況作為卡那霉素處理效果的篩選標準,即作為非轉基因陰性株。
       2.2 轉基因抗蟲品種的棉花葉片對卡那霉素的反應效果
      隨著卡那霉素溶液濃度的提高,轉基因抗蟲品種抗蟲5號、抗蟲10號的棉花葉片,未出現明顯變化(表1)。說明適當提高卡那霉素溶液濃度,在提高卡那霉素溶液對轉基因抗蟲棉花檢測準確率的同時,不會出現誤檢現象。
       2.3 不同品種的非轉基因棉花葉片對卡那霉素的反應效果
      選擇3個有代表性的非轉基因棉花品種和2個轉基因抗蟲棉品種作為供試材料,研究不同棉花品種對卡那霉素處理的外觀表現差異。結果表明:棉花不同品種間葉片對卡那霉素處理的外觀表現差異不明顯,說明不同的棉花品種對卡那霉素處理外觀變化相同,因此用卡那霉素篩選、鑒定轉基因抗蟲棉花品種,適用性很大。
       2.4 棉花葉片對不同濃度的卡那霉素溶液的反應效果
      在試驗過程中發現(表1),隨著卡那霉素濃度的提高,各個試驗處理中的假陽性植株比例逐漸減小。當卡那霉素溶液濃度達到5 000 mg/l時,檢測的準確度最好。卡那霉素對棉花葉片處理效果的受諸多因素影響,其中之一為棉花葉片對卡那霉素的抗性,在棉花的抗性范圍內,棉花葉片個體間的外觀表現差異較大,而當卡那霉素濃度超出抗性范圍后,棉花葉片個體之間的外觀表現趨于一致。
       3 討論
      卡那霉素對作物細胞表現毒性的機理是:干擾細胞葉綠體、線粒體的蛋白質合成,使植物細胞死亡。轉基因植株抑制這個干擾過程的進行,因此通過卡那霉素處理棉花葉片篩選、鑒定轉基因棉花植株是具有理論依據的。卡那霉素是目前基因工程中廣泛使用的選擇標記,因此在棉花的生育早期進行卡那霉素檢測,適當提高卡那霉素處理濃度,得到的假陽性單株少;轉基因篩選、鑒定的效果較好。經過不同濃度處理的篩選效果對比,當卡那霉素溶液濃度達到5 000 mg/l時,檢測的準確度最好。
       4 參考文獻
      [1] 祝建波,王愛英,封玲.棉花葉片對卡那霉素反應效果的研究[j].石河子大學學報,1999,3(3):187-191.
      [2] 葛朝紅,孟建朝,田海燕.卡那霉素對棉花不同時期抗蟲性的鑒定[j].現代農村科技,2009(16):10-11.
      [3] 李虎申,張岳華,李莉,等.轉基因抗蟲棉純度的幼苗鑒定方法研究[j].中國種業,2007(3):47-48.
      [4] 李秀珍,儲甲松,彭華升,等.轉基因棉花在生產應用上的風險與安全控制[j].江西棉花,2010,32(4):47-49.

     

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