不同地理种源黑果枸杞的组织培养研究

发布时间：2018-03-09 13:09

本文选题：黑果枸杞　切入点：不同种源　出处：《甘肃农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：黑果枸杞(Lycium barbarum),系茄科(Solanaceae)枸杞属(Lycium L.)植物,是我国西北干旱地区特有的兼具耐盐碱和抗旱特性的野生灌木,可正常生长于大多植物都无法生长的高盐渍化环境中。近年来,因其丰富的营养价值、商用价值及潜在的药用价值,加上经济利益的驱动,野生资源受到大规模的掠夺性开采,致使黑果枸杞天然分布区自然条件日益恶劣,植被严重退化甚至死亡,已经对其它伴生植物的的生长造成很大的威胁,野生居群的规模以及分布区域呈迅速减小的趋势。然而野生资源由于数量少,远远不能满足市场的需求,而利用人工播种繁殖易出现遗传分化、变异,扦插育苗繁殖系数低的缺点,大大影响了果实的品质。因此,对其组织培养和快速繁殖显得尤为重要,为满足黑果枸杞种质资源保护及商品化及规模化生产、推广应用提供了理论依据。本研究以我国新疆、青海、甘肃民勤三个不同地理种源的黑果枸杞为试验材料,运用组织培养技术对这三种植物进行快速繁殖。主要研究结果如下:(1)三个不同种源黑果枸杞的不同外植体最佳灭菌时间因种源和外植体取材部位的不同而不同。新疆黑果枸杞叶片最佳灭菌时间为75%乙醇10s、HgCI23min,茎段最佳灭菌时间为75%乙醇10s、HgCI25min;青海黑果枸杞叶片最佳灭菌时间为75%乙醇10s、HgCI2 3min,茎段为75%乙醇20s、HgCI23min;民勤黑果枸杞叶片最佳灭菌时间为75%乙醇10s、Hg CI23min,茎段最佳灭菌时间为75%乙醇20s、HgCI2 5min。(2)最适宜新疆黑果枸杞进行愈伤组织培养的培养基配方为:MS+6-BA1.5mg/L+2,4-D0.05mg/L+蔗糖30g/L+琼脂5g/L;MS+6-BA1.0mg/L+2,4-D 0.1mg/L+蔗糖30g/L+琼脂5g/L是青海黑果枸杞的最佳培养基配方;民勤地区黑果枸杞在MS+6-BA1.0mg/L+2,4-D 0.1mg/L+蔗糖30g/L+琼脂5g/L中培养的效果最好。(3)MS+NAA0.5 mg/L+6-BA1.0 mg/L+蔗糖30g/L+琼脂5g/L是新疆黑果枸杞最适不定芽分化培养基配方;青海黑果枸杞在MS+NAA0.5mg/L+6-BA0.5mg/L+蔗糖30g/L+琼脂5g/L培养基中不定芽分化效果最理想;民勤黑果枸杞的最佳不定芽分化培养基配方为MS+NAA0.5mg/L+6-BA0.5mg/L+蔗糖30g/L+琼脂5g/L。(4)MS+NAA0.5mg/L+6-BA0.1mg/L+蔗糖30g/L+琼脂5g/L是新疆黑果枸杞最适的继代增殖培养基配方;青海黑果枸杞在MS+NAA0.5mg/L+6-BA0.3mg/L+蔗糖30g/L+琼脂5g/L中继代增殖效果最佳;民勤黑果枸杞的最佳继代增殖配方为MS+NAA0.5 mg/L+6-BA0.1 mg/L+蔗糖30g/L+琼脂5g/L。(5)新疆黑果枸杞在1/2MS+IBA0.1mg/L+蔗糖30g/L+琼脂5g/L培养基中生根与生长效果最佳;青海黑果枸杞最适生根培养基配方为:l/2MS+IBA0.1mg/L+蔗糖10g/L+琼脂5g/L,1/2MS+蔗糖30g/L+琼脂5g/L是民勤黑果枸杞最佳的生根培养基配方。研究表明三个不同地理种源的黑果枸杞均适合于使用组织培养的方法进行快速繁殖,当植物生长调节物质以适宜的浓度组合配比时,三个种源的黑果枸杞均可在4-5个月内再生出生长、发育健壮的试管苗,完全可在大规模工厂化生产进行应用。
[Abstract]:Lycium barbarumum (Solanaceaeaceae) Lycium L.) is a wild shrub with both saline-alkali and drought-resistant characteristics, which is endemic to arid areas of northwest China. It can normally grow in a highly salinized environment where most plants cannot grow. Because of its rich nutritional value, commercial value and potential medicinal value, and driven by economic benefits, wild resources have been exploited on a large scale, resulting in the worsening of natural conditions in the natural distribution area of Black Fruit and Lycium barbarum. Serious degradation or even death of vegetation has posed a great threat to the growth of other associated plants, and the size and distribution of wild populations have tended to decrease rapidly. However, due to the small number of wild resources, It is far from meeting the demand of the market, but it is easy to produce genetic differentiation, variation and low propagation coefficient of cuttage breeding by artificial seeding, which greatly affects the quality of fruit. Therefore, it is very important for its tissue culture and rapid propagation. In order to protect the germplasm resources of black fruit wolfberry and produce it on a commercial scale and in a large scale, a theoretical basis was provided for its popularization and application. Three different provenances of Xinjiang, Qinghai and Minqin in Gansu Province were used as experimental materials in this study. The main results are as follows: (1) the optimum sterilization time of different explants of three different provenances of Lycium barbarum is different with different provenances and explants. The optimum sterilizing time of black fruit Lycium barbarum leaves in Xinjiang is 75% ethanol 10sL HgCI 23min, the optimum sterilizing time of stem segment is 75% ethanol 10sL HgCI 25min, the optimum sterilization time of Qinghai black fruit Lycium barbarum leaves is 75% ethanol 10sL HgCI23mins, the stem segment is 75% ethanol 20sHgCI23mins; the Minqin black fruit Lycium barbarum leaf is best to kill. The optimum medium for callus culture of Xinjiang black fruit wolfberry was: 1: MS 6-BA 1.5mg / L 4-D 0.05mg / L sucrose 30g / L Agar 5g / L MS 6-BA1.0mg / L 0.1 mg / L sucrose 5g / L Agar 5g / L Agar 5g / L MS 6-BA1.0mg / L 0.1 mg / L sucrose 5g / L. The best medium formulation of Lycium barbarum; The best culture effect of Chinese wolfberry in MS 6-BA 1.0mg / L 24-D 0.1 mg / L sucrose 30g / L Agar 5g / L was found to be the most suitable medium for adventitious bud differentiation in Minqin area. Ms NAA0.5 mg/L 6-BA1.0mg / L sucrose 30g / L sucrose 30g / L Agar 5g / L was the most suitable medium for adventitious bud differentiation of Chinese wolfberry. In MS NAA0.5mg/L 6-BA 0.5mg / L sucrose 30g / L Agar 5g / L medium, the best differentiation effect of adventitious buds was obtained. The best medium for adventitious bud differentiation was MS NAA0.5mg/L 6-BA0.5mg / L sucrose 30g / L Agar 5g / L Agar 5g / L MS NAA0.5mg/L 6-BA0.1mg / L sucrose 30g / L Agar 5g / L was the best subculture medium for Xinjiang black fruit wolfberry. In MS NAA0.5mg/L 6-BA 0.3 mg / L sucrose 30 g / L Agar 5g / L relay generation, the best proliferative effect of Qinghai Black Fruit Lycium barbarum L. The best subculture multiplication formula was MS NAA0.5 mg/L 6-BA0.1 mg/L sucrose 30g / L Agar 5g / L Agar 5g / L). Xinjiang Black Fruit Lycium barbarum was rooting and growing best in 1 / 2ms IBA0.1mg/L sucrose 30g / L Agar 5g / L medium. The optimum rooting medium of Qinghai black fruit wolfberry is: 1 / 1 / 2ms IBA0.1mg/L sucrose 10 g / L Agar 1 / 2 MS sucrose 30 g / L Agar 5g / L Agar 5g / L is the best rooting medium for Minqin black fruit medlar. Rapid propagation by tissue culture, When the plant growth regulator was combined with suitable concentration, the three provenances of Lycium barbarum could regenerate in 4-5 months and develop robust plantlets, which could be used in large-scale industrial production.
【学位授予单位】：甘肃农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S567.19

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