花生幼叶丛生芽高效诱导的制约因素的研究.docVIP

花生幼叶丛生芽高效诱导的制约因素的研究 摘要:以花生品种豫花14号4d苗龄的幼叶为外植体,对花生幼叶高频不定芽诱导进行研究在MS培养基的基础上,添加了芽诱导常用的细胞分裂素6-BA、生长素NAA及两种不太常用的脱落酸(ABA)、噻重氮苯基脲(TDZ),结果表明,单独使用6-BA,NAA或TDZ,不定芽诱导率较低;采用较高浓度的6-BA(8mg/L)、较低浓度的NAA(1mg/L),添加低浓度的ABA(1mg/L),不定芽诱导率可达80%以上。最佳诱导培养基为MS+6-BA 8mg/L+NAA 1mg/L+ABA 1mg/L +AgNO32mg/L,从整体上观察,最佳继代培养基为MS+6-BA 5mg/L+NAA 2mg/L+AgNO32mg/L。同时研究发现,花生幼叶近叶柄基部切口处不定芽诱导率较高,是较理想的不定芽诱导部位。 关键词:花生;组织培养;不定芽诱导;植株再生. The Restrictive Factors for Efficient Induction ofAdventitious Buds from Peanut Leafletin vitro Abstract: The efficiency of adventitious bud induction from leaflet were investigated using the 4- day seedlings of peanut variety Yuhua 14.The MS medium was added with commonly-used kinin 6-BA,kinetin NAA,and uncommonly-used ABA and thiadiazole Diazo phenyl urea TDZ.Results showed that the adventitious bud induction rate was low in the MS added with only 6-BA,NAA and TDZ,but reached to 80% in the MS added with high concentrations of 6-BA (8mg/L),low concentrations of NAA (1mg/L) and low concentrations ABA (1mg/L),indicating that the MS medium+8mg/L 6-BA+1mg/L NAA+1mg/L ABA+2mg/L AgNO3had the best result for inducing the adventitious buds.The best medium for subculture was MS+5mg/L 6-BA+2mg/ L NAA+2mg/L AgNO3.It was also observed that the underside cut near the petiole presented a higher rate of adventitious buds,so was the ideological explant for adventitious bud induction. Key words: Peanut; Tissue culture; Adventitious buds induction; Regeneration 　　花生是重要的油料和经济作物,在世界油脂生产中占有举足轻重的地位。随着我国人民生活水平的提高、种植业结构的调整和对外贸易的扩大,根据不同的市场需要,农业生产上对花生品种的需求也向多元化和专用型发展,选育不同脂肪酸含量和比例的加工型(高硬脂酸含量)、保健型(高多不饱和脂肪酸)、出口型(高O/L比)花生新种质是花生新品种选育的重要目标[1]。实践证明,利用基因工程技术进行油料作物脂肪酸含量和组分调控是种质创新的有效途径[2~4]。花生组织培养及高效植株再生体系的建立是基因转化及外源基因利用的基础,但目前花生的组培植株再生体系不够理想,成为基因转化的限制因素之一[5]。为此,选用河南省大面积推广的花生品种豫花14号的幼叶作外植体,采用不同激素配比的诱导培养基进行了高效丛生芽诱导和植株再生体系研究,旨在为进一步研究基因转化和脂肪酸调控奠定基础。 1.　材料 1.1　植物材料　豫花14号花生品种,亚油酸含量42%,系河南省农业科学院经济作物研究所培育的高亚油酸品种,由河南省农业科学院经济作物研究所提供。 1.2　外植体　分别采用萌发4d的幼叶为外植体。 1.3　植物激素　N-苯基-N′-1,2,3-噻二唑-5-脲(噻重氮苯基脲,TDZ);6-苄基嘌呤(6- BA);a-萘乙酸(N