华北农学报
華北農學報
화북농학보
ACTA AGRICULTURAE BOREALI-SINICA
2009年
5期
35-39
,共5页
黄亚丽%蒋细良%田云龙%朱昌雄
黃亞麗%蔣細良%田雲龍%硃昌雄
황아려%장세량%전운룡%주창웅
哈茨木霉%T-DNA%突变子%TAIL-PCR%厚垣孢子
哈茨木黴%T-DNA%突變子%TAIL-PCR%厚垣孢子
합자목매%T-DNA%돌변자%TAIL-PCR%후원포자
Trichoderma harzianum%T-DNA%Mutant%TAIL-PCR%Chlamydospore
利用PD液体培养基从哈茨木霉T-DNA插入突变体库中筛选出在产孢性状上与野生型菌株明显不同突变子7株,其突变表型主要表现在分生孢子产生数量显著减少和菌丝上有大量厚垣孢子分化.利用TAIL-PCR方法对7株突变体的侧翼序列进行克隆,从5个突变体中获取了5条T-DNA侧翼序列.为木霉菌厚垣孢子产生相关全长基因的克隆和产孢机制的研究奠定了基础.
利用PD液體培養基從哈茨木黴T-DNA插入突變體庫中篩選齣在產孢性狀上與野生型菌株明顯不同突變子7株,其突變錶型主要錶現在分生孢子產生數量顯著減少和菌絲上有大量厚垣孢子分化.利用TAIL-PCR方法對7株突變體的側翼序列進行剋隆,從5箇突變體中穫取瞭5條T-DNA側翼序列.為木黴菌厚垣孢子產生相關全長基因的剋隆和產孢機製的研究奠定瞭基礎.
이용PD액체배양기종합자목매T-DNA삽입돌변체고중사선출재산포성상상여야생형균주명현불동돌변자7주,기돌변표형주요표현재분생포자산생수량현저감소화균사상유대량후원포자분화.이용TAIL-PCR방법대7주돌변체적측익서렬진행극륭,종5개돌변체중획취료5조T-DNA측익서렬.위목매균후원포자산생상관전장기인적극륭화산포궤제적연구전정료기출.
7 mutants with chlamydospore formation differences were selected from 1 192 transformants cultured in PD liquid medium.The conidia formation ability of all these 7 mutants was significantly lower than that of the wild type T. harzianum .The characterization of these 7 mutants was analyzed.The results showed that as for the mutants 918,1 137, 1 317 chlamydospore developed in the mycelium and failed off from mycelium when incubated time prolonged .However, as for the other 4 mutants 627,840,1 137,1 173 the chlamydospore couldn't failed off from mycelium when incubated time prolonged. Using TAIL-PCR to clone T-DNA right border sequences from spore formation mutants, we successfully obtained the fungal genomic DNA sequences flanking T-DNA right border from 5 transformants. The homologous sequences of 3 sequences were found by using Blastn and Blastx from NCBI GenBank. However, the homologous sequences of other 2 sequences were not found. They might be new genes related to spore formation which need to be further studied. This study is the base for the cloning of chlamydospore related gene and the mechanism research of chlamydospore formation.
