發(fā)布時間：2019-04-09 11:37

【摘要】：在前期工作中,將Bt cry1Ah基因轉(zhuǎn)入玉米自交系綜31中,選取其中一株對亞洲玉米螟有顯著抗性的玉米植株,命名為HGK60。分別從DNA水平和轉(zhuǎn)錄翻譯水平對HGK60事件的T4代和T5代植株進行遺傳穩(wěn)定性分析。通過PCR、Tail-PCR、Southern Blot分析表明外源基因整合到了玉米基因組中并可以穩(wěn)定遺傳給后代,且明確了外源基因是按單插入雙拷貝的方式插入到玉米1號染色體上,分析得到了玉米插入序列的側(cè)翼序列。通過RT-PCR、Western Blot、ELISA、試紙條分析表明外源基因可以在玉米的根、莖、葉等組織中正常高效表達。在室內(nèi)和田間分別用亞洲玉米螟、棉鈴蟲和黏蟲幼蟲對HGK60玉米的殺蟲效果進行檢測。室內(nèi)生物活性檢測結(jié)果表明,取食HGK60玉米心葉的亞洲玉米螟幼蟲在3天后死亡率達100%;HGK60玉米對棉鈴蟲幼蟲有毒殺作用,玉米不同部位對棉鈴蟲幼蟲的殺蟲效果存在差異;與綜31相比,取食HGK60玉米葉片一周后的黏蟲幼蟲體重增長明顯受到抑制。田間生物活性檢測結(jié)果表明,HGK60玉米對于亞洲玉米螟和棉鈴蟲有很強的殺蟲活性,達到高抗級別,對于黏蟲有一定的殺蟲活性,為抗性級別。對HGK60與鄭58自交系回交轉(zhuǎn)育5代后的轉(zhuǎn)基因玉米進行農(nóng)藝性狀分析。在無蟲害情況下,與鄭58相比,HGK60的株高、穗位高、穗粗、穗長等差異不顯著;在人工模擬蟲害爆發(fā)的的情況下,鄭58的生長發(fā)育受到影響,千粒重和單株產(chǎn)量都顯著低于HGK60。以上研究表明,HGK60轉(zhuǎn)基因抗蟲玉米在玉米的抗蟲育種上有很好的應(yīng)用前景。
[Abstract]:In previous work, Bt cry1Ah gene was introduced into maize inbred ensemble 31, and one of the maize plants with significant resistance to Asian corn borer, named HGK60., was selected. The genetic stability of T4 and T5 generations of HGK60 events was analyzed at DNA level and transcriptional translation level, respectively. PCR,Tail-PCR,Southern Blot analysis showed that the exogenous gene was integrated into the maize genome and could be stably inherited to the progeny, and it was clear that the exogenous gene was inserted into maize chromosome 1 in the form of single insertion and double copy. The flanking sequence of maize insertion sequence was analyzed. The results of RT-PCR,Western Blot,ELISA, analysis showed that exogenous genes could be expressed normally and efficiently in roots, stems, leaves and other tissues of maize. The insecticidal effects of Asian corn borer, Helicoverpa armigera and Myxworm larvae on HGK60 maize were tested in laboratory and field, respectively. The results of laboratory bioactivity test showed that the mortality of Asian corn borer larvae fed on the heart leaves of HGK60 corn was 100% HGK60 after 3 days, and the insecticidal effects of different parts of corn on the larvae of cotton bollworm were different. Compared with Heizang 31, the weight gain of myxozoa larvae after feeding HGK60 maize leaves for one week was significantly inhibited. Field bioassay showed that HGK60 maize had strong insecticidal activity against Asian corn borer and Helicoverpa armigera, which reached high resistance level, and had certain insecticidal activity to Myxworm, which was the resistance level of corn borer and Helicoverpa armigera. The agronomic characters of transgenic maize after 5 generations of backcross between HGK60 and Zheng 58 inbred line were analyzed. Compared with Zheng 58, there was no significant difference in plant height, ear position, ear diameter and ear length between HGK60 and Zheng 58. Under the condition of artificial pest outbreak, the growth and development of Zheng 58 were affected, and the 1000-grain weight and yield per plant were significantly lower than that of HGK60.. The results indicated that HGK60 transgenic insect-resistant maize had good application prospect in maize pest-resistant breeding.
【學(xué)位授予單位】：西南科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S513
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本文編號：2455147