摘要
成熟子叶切块作为外殖体,经过组织培养获得再生植株。以MS作为基本培养基,取3日龄子叶块外殖体,选用MS+6-BA1.5mg·L-1+IAA0.3mg·L-1诱导从生芽,最高诱导率可达78%。分化再生过程中,6-BA浓度逐渐降低。在生根培养时,用MS+0.3mg·L-1NAA诱导生根率较高,根系粗壮。在建立甜瓜高效再生体系的基础上,用根癌农杆菌介导法将几丁质酶基因和β-1,3-葡聚糖酶基因导入新疆甜瓜中,经卡钠霉素的抗性筛选,获得转化的再生植株。通过PCR反应及Southern Blot检测,进一步研究了再生植株的基因整合与表达情况,获得与阳性对照一致的特异带,可初步证明外源基因已整合到甜瓜基因组中。
In order to establish the transgenic plant system, the cotyledons of muskmelon as explants, regenerated plantlets were used in culture. Shoots regeneration were induced on MS medium as basic medium supplemented with 1.5 mg·L^-1 6-BA＋0.3 mg·L^-1 IAA. The highest frequency shoot regeneration observed was 78%. To achieve the high rate of rooting, the concentration of hormones 6-BA was gradually decreased during the whole growth period. The 1/2 MS medium with 0.3 mg·L^-1NAA was used for this propose. On the basis of regeneration protocol of Xinjiang muskmelon, two genes were transferred into muskmelon plant and selected by kanamycin in the medium. After kanamycin resistant plants were obtained, integration of these genes into the genome of muskmelon plant was confirmed by PCR, Southern Blot analyses.
出处
《东北农业大学学报》
CAS
CSCD
2008年第1期67-70,共4页
Journal of Northeast Agricultural University
基金
新疆新天石大科学基金项目(XS200009)
