黑果枸杞优质种苗快速繁殖的方法 Black wolfberry fruit quality seeds and seedlings rapid propagation method

技术领域 Technical Field

[0001] 本发明涉及一种黑果枸杞培育方法，更具体地说涉及一种黑果枸杞优质种苗快速繁殖的方法。 [0001] The present invention relates to a method of cultivating black fruit wolfberry, and more particularly to a high-quality black fruit wolfberry seedlings rapid propagation method.

背景技术 Background

[0002] 黑果JMHiZycium ruthenicum Murr.)为爺科枸杞,属于一种盐生药用植物,在我国主要分布在宁夏、山西、甘肃、青海、新疆、西藏等省区。 [0002] black fruit JMHiZycium ruthenicum Murr.) For the Lord Section wolfberry, is a salt raw medicinal plant in China is mainly distributed in Ningxia, Shanxi, Gansu, Qinghai, Xinjiang, Tibet and other provinces. 黑果枸杞用途广泛，其果实中含有多种人体必需的氨基酸和微量元素，可提高人体免疫能力；从果汁中提取的黑果枸杞色素稳定性好、着色力强，是国内急需的理想食用天然花色甙，不但可应用于药品和食品中，而且可作为天然染料应用于轻纺工业，是迄今为止发现原花青素含量最高的野生植物。 Black Murr versatile, its fruit contains a variety of essential amino acids and trace elements can improve the immune capacity; good PIGMENT extracted from the juice stability, strong coloring, is the urgent need of the ideal edible natural Anthocyanin, not only can be used in medicines and foods, and can be used as a natural dye used in textile industry, is by far the highest concentrations found in wild plants proanthocyanidins. 同时，黑果枸杞具有抗旱、抗寒、耐盐碱、根蘖性强和耐土壤贫瘠等诸多优点，是盐碱地治理、护坡、防止水土流失的良好树种。 Meanwhile, the black fruit wolfberry drought, cold, salinity, strong and resistant root suckers poor soils and many other advantages, is saline control, slope protection, prevent soil erosion of good species. 因此，黑果枸杞无论在经济产品开发，还是在生态环境治理方面都具有广泛的市场前景和开发潜力，可以成为医药保健、经济林建设和盐碱地生物防治的重要经济树种。 Therefore, black fruit wolfberry products both in economic development, or in terms of the ecological environment have broad market prospects and development potential, can become health care, construction and saline forest biological control important economic species. 目前，黑果枸杞的研究绝大多数集中在对其生理生态、有效成分分析、开发利用以及治理荒漠化和野生种源驯化等方面，而关于黑果枸杞组织培养与快速繁殖方面的研究报到很少。 At present, the vast majority of black fruit wolfberry concentrated in their physiological ecology, the active ingredient analysis, exploitation and control of desertification and other aspects of domestication of wild provenance, and research about black fruit wolfberry tissue culture and rapid breeding of the rare report . 浩仁塔本等(2005)利用带腋芽的茎段对黑果枸杞进行了分化生根及愈伤组织诱导不定芽增殖进行了研究，而有关黑果枸杞优质种苗快速繁殖方面的研究还未见报道。 Hao Ren tower of the (2005) the use of stem with axillary bud of Black Murr were rooting and callus induction of differentiation of adventitious buds were studied, and the study of black fruit wolfberry rapid breeding of quality seeds and seedlings have not been reported . 目前甘肃省黑果枸杞野生资源主要分布区自然条件恶劣，植被退化严重，甚至成片死亡，已对黑果枸杞的正常生存造成了严重威胁，基本处于濒危状态。 Currently, Gansu Province wild black fruit wolfberry main distribution area resources harsh natural conditions, vegetation degradation is serious, and even death into a film, has black fruit of Chinese wolfberry a serious threat to the survival of the normal, basic endangered.

发明内容 DISCLOSURE

[0003] 本发明要解决的技术问题是:解决我省黑果枸杞品种选育和良种繁殖的问题，解决种子繁殖周期长的问题，提供一种黑果枸杞优质种苗快速繁殖的方法，获得优质种苗，缩短黑果枸杞繁殖周期，加快黑果枸杞良种繁育进程，可进行工厂化大规模生产，满足市场对黑果枸杞的需求，保护野生黑果枸杞自然资源和良好的自然生态环境。 [0003] The technical problem to be solved is: solve the province black wolfberry fruit breeding and seed breeding problems, solving a long period of seed multiplication problems and provide a high-quality black fruit wolfberry seedlings rapid propagation methods to obtain quality seeds and seedlings, shorten the breeding cycle of black wolfberry fruit, black fruit wolfberry accelerate breeding process can be carried out large-scale factory production to meet the market demand for black fruit wolfberry, protection of wild black fruit wolfberry natural resources and a healthy natural environment.

[0004] 本发明通过下述顺序步骤实现的: [0004] The present invention is achieved by the following sequence of steps:

a、首先选择黑果枸杞优质种子用流水冲洗2〜3h ; a, first select black fruit wolfberry quality seeds with running water 2~3h;

b、将冲洗后的种子用75%酒精消毒lmin，然后将酒精消毒后的种子用无菌水冲洗3〜4次； b, the seeds rinsed with 75% alcohol disinfection lmin, then rinse 3 to 4 times the seed of alcohol after disinfection with sterile water;

C、将无菌水冲洗后的种子再用0.1%的升汞消毒5min，然后将升汞消毒后的种子用无菌水冲洗3〜4次； C, the seeds sterile water and then after 0.1% mercuric chloride for 5min, then the seeds of mercuric chloride after 3 to 4 times with sterile water;

d、最后将经过灭菌处理的种子接种于MS培养基中培养无菌苗，培养温度为24°C〜26°C，光照强度为1500 LX〜2000LX，光照时间为24h/d ； d, and finally sterilized through seed inoculated in MS culture medium without vaccine, the culture temperature was 24 ° C~26 ° C, light intensity of 1500 LX~2000LX, illumination time 24h / d;

e、培养I个月后，将无菌苗的茎杆剪下，切成0.5cm长短的茎段，于无菌条件下接入MS+KH2PO4 13.5mg/L +NAA 0.lmg/L培养基中进行无菌苗的快速扩繁培养，光照强度为2000LX，光照时间为12h/d〜16h/d。 e, I cultivate months, there will be no vaccine stems cut, cut into 0.5cm length of the stem, under sterile conditions, access MS + KH2PO4 13.5mg / L + NAA 0.lmg / L medium carried aseptic culture fast propagation, light intensity 2000LX, illumination time 12h / d~16h / d. [0005] 本发明利用组织培养方法快速繁殖黑果枸杞无菌苗的茎杆，其优点是:使无菌苗生根率达100%，可快速繁殖黑果枸杞无菌苗木。 [0005] The present invention utilizes the tissue culture and rapid propagation of black fruit wolfberry aseptic stem, its advantages are: so that no vaccine is 100% rooting rate, rapid propagation of black fruit wolfberry sterile seedlings. 周期短，不受季节、气候、自然灾害等因素的影响，并可实现工厂化生产。 Short period, is not affected by seasonal factors, weather, natural disasters, and to achieve industrial production. 利用这种技术可以解决药用植物的繁殖能力低、资源匮乏、农药残留等诸多问题。 Using this technology can solve the low reproductive capacity of medicinal plants, lack of resources, pesticide residues and many other issues. 另一方面我们甘肃是首次采用植物组织培养技术，这对甘肃黑果枸杞新种质资源进行快速繁殖，不仅可以满足市场需求，而且还在防风固沙、保护生态环境和保护种质资源等方面具有战略意义。 Gansu is the first time the other hand we of plant tissue culture techniques, which Gansu black fruit wolfberry rapid propagation of new germplasm resources, not only to meet the market demand, but also wind and sand, protecting the environment and the protection of germplasm resources have strategic significance. 具体实施方式[0006] 实施例1 本发明有下述顺序步骤: a、首先选择黑果枸杞优质种子用流水冲洗2〜3h ; b、将冲洗后的种子用75%酒精消毒lmin，然后将酒精消毒后的种子用无菌水冲洗3〜4次； C、将无菌水冲洗后的种子再用0.1%的升汞消毒5min，然后将升汞消毒后的种子用无菌水冲洗3〜4次； d、最后将经过灭菌处理的种子接种于MS培养基中培养无菌苗，培养温度为24°C，光照强度为1500 LX，光照时间为24h/d ； e、培养I个月后，将无菌苗的茎杆剪下，切成0.5cm长短的茎段，于无菌条件下接入MS+ KH2PO4 13.5mg/L +NAA 0.lmg/L培养基中继续进行无菌苗的快速扩繁培养，光照强度为2000LX，光照时间为12h/d。 DETAILED DESCRIPTION [0006] Example 1 of the present invention has the following sequential steps: a, first select the black fruit wolfberry quality seeds with running water 2~3h; b, the seeds rinsed with 75% alcohol disinfection lmin, and then the alcohol After disinfection rinse the seeds 3 to 4 times with sterile water; C, the seeds sterile water and then after 0.1% mercuric chloride for 5min, then sterilized seeds of mercuric chloride rinsed with sterile water 3 to 4 times; d, and finally sterilized through seed inoculated in MS culture medium without vaccine, the culture temperature was 24 ° C, light intensity of 1500 LX, illumination time 24h / d; e, I cultivate months , there will be no vaccine stems cut, cut into 0.5cm length of the stem, under sterile conditions, access MS + KH2PO4 13.5mg / L + NAA 0.lmg / L medium continued rapid aseptic culture propagation, light intensity 2000LX, illumination time 12h / d. [0007] 实施例2 实施例2与实施例1的方法相同，所不同的是步骤d中无菌苗的培养温度为25°C，光照强度为1800LX。 [0007] Example 2 Example 2 in the same manner as in Example 1, except that in step d in aseptic culture temperature of 25 ° C, light intensity 1800LX. 步骤e中光照时间为14h/d。 Step e illumination time 14h / d. [0008] 实施例3 实施例3与实施例1的方法相同，所不同的是步骤d中无菌苗的培养温度为26°C，光照强度为2000LX。 [0008] Example 3 Example 3 in the same manner as in Example 1, except that in step d in aseptic culture temperature of 26 ° C, light intensity 2000LX. 步骤e中光照时间为16h/d。 Step e illumination time 16h / d. [0009] 综上所述:该项技术使无菌苗生根率达100%，可快速繁殖黑果枸杞无菌苗木。 [0009] To sum up: the technology so that no vaccine is 100% rooting rate, rapid propagation of black fruit wolfberry sterile seedlings. 甘肃是首次采用黑果枸杞无菌苗茎杆快速扩繁技术，这对甘肃黑果枸杞新种质资源进行快速繁殖，不仅可以满足市场需求，而且还在防风固沙、保护生态环境和保护种质等方面具有战略意义。 Gansu is the first time black fruit wolfberry aseptic stem rapid propagation technology, which Gansu black fruit wolfberry rapid propagation of new germplasm resources, not only to meet the market demand, but also wind and sand, protecting the environment and protecting the germplasm so has a strategic significance.