杜仲带芽茎段诱导和分化的组培体系优化
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摘要
以杜仲的幼嫩茎段为试材,采用组织培养的方法,研究了消毒时间、基本培养基、NAA浓度、6-BA浓度、低温处理时间及培养环境对杜仲腋芽诱导及其分化的影响,以期为建立杜仲高效组织培养体系提供依据。结果表明:最佳消毒时间为9 min,污染率与死亡率均较低,分别为10.00%和26.67%;腋芽诱导的基本培养基以MS培养基为最佳,诱导率达91.66%;诱导培养基为MS+0.2 mg·L~(-1) 6-BA+0.03 mg·L~(-1) NAA时,腋芽的诱导率较高,达93.33%,且生长良好;当低温处理时间为24 h时,外植体诱导效果最好,且褐化率最低,为5.00%;与黑暗培养相比,光照培养条件更加有利于杜仲的腋芽诱导和生长;MS+1.5 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA为愈伤分化的最佳培养基,MS+2.5 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA为芽增殖的最佳培养基。
The young stem segments of Eucommia ulmoides Oliver were used as materials,tissue culture method was used,the effects of disinfection time,basic medium,NAA concentration,6-BA concentration,low temperature treatment time and culture environment on the induction of Eucommia ulmoides Oliver axillary buds and the differentiation of Eucommia ulmoides Oliver were investigated,in order to provide a basis for establishing an efficient tissue culture system of Eucommia ulmoides Oliver.The results showed that the best disinfection time was 9 minutes,and the rate of pollution and mortality were lower,10.00% and 26.67% respectively;the basic medium for axillary buds induction was MS medium,with an induction rate of 91.66%;when the induction medium was MS+0.2 mg·L~(-1) 6-BA+0.03 mg·L~(-1) NAA,the induction rate of axillary buds was 93.33% and it grew well;when the low temperature treatment time was 24 hours,explant induction effect was the best,and browning rate was the lowest,5.00%;compared with dark culture,light culture conditions were more conducive to the induction and growth of axillary buds of Eucommia ulmoides;MS+1.5 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA was the best medium for callus differentiation,MS+2.5 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA was the best medium for bud proliferation.
The young stem segments of Eucommia ulmoides Oliver were used as materials,tissue culture method was used,the effects of disinfection time,basic medium,NAA concentration,6-BA concentration,low temperature treatment time and culture environment on the induction of Eucommia ulmoides Oliver axillary buds and the differentiation of Eucommia ulmoides Oliver were investigated,in order to provide a basis for establishing an efficient tissue culture system of Eucommia ulmoides Oliver.The results showed that the best disinfection time was 9 minutes,and the rate of pollution and mortality were lower,10.00% and 26.67% respectively;the basic medium for axillary buds induction was MS medium,with an induction rate of 91.66%;when the induction medium was MS+0.2 mg·L~(-1) 6-BA+0.03 mg·L~(-1) NAA,the induction rate of axillary buds was 93.33% and it grew well;when the low temperature treatment time was 24 hours,explant induction effect was the best,and browning rate was the lowest,5.00%;compared with dark culture,light culture conditions were more conducive to the induction and growth of axillary buds of Eucommia ulmoides;MS+1.5 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA was the best medium for callus differentiation,MS+2.5 mg·L~(-1) 6-BA+0.1 mg·L~(-1) NAA was the best medium for bud proliferation.
引文
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[15] 德永军,吕涛,王一,等.文冠果茎段形成层不定芽诱导组织培养技术初探[J].内蒙古农业大学学报(自然科学版),2014,35(2):39-42.
[16] 张桂琴,徐祥龄,赵志学.文冠果嫩茎组织诱导植株移栽初获成功[J].林业科技通讯,1980(7):4-5.
[17] 张娜,张芸香,郭晋平.文冠果腋芽诱导关键影响因素与培养体系优化[J].山西农业大学学报(自然科学版),2014,34(1):53-58.
[18] 李琰,张存莉,严忠海,等.不同培养基对杜仲愈伤组织诱导和生长的影响[J].西北林学院学报,2003(3):37-39.
[19] 高国训.植物组织培养中的褐变问题[J].植物生理学通讯,1999(6):501-506.
[20] 黄烈健,王鸿.林木植物组织培养及存在问题的研究进展[J].林业科学研究,2016,29(3):464-471.
[21] 邢阿宝,崔海峰,俞晓平,等.光质及光周期对植物生长发育的影响[J].北方园艺,2018(3):163-172.
[22] SKOOG F,MILLER C O.Chemical regulation of growth and organ formation in plant tissues cultured in vitro[J].Symp Soc Exp Biol,1957,54:118-130.
[23] 谷文英,裴德清,朱登云,等.杜仲茎段培养的初步研究[J].莱阳农学院学报,1999(1):6-9.
[2] 康向阳.杜仲良种选育研究现状及展望[J].北京林业大学学报,2017,39(3):1-6.
[3] 童孝茂.杜仲的栽培技术和开发利用[J].安徽农学通报,2007(20):117-118.
[4] 朱翠英,张真,谢经霞,等.杜仲组培快繁技术的研究[J].宁夏农林科技,2005(6):32-33.
[5] 张康健.中国杜仲研究[M].西安:陕西科技出版社,1992:21
[6] LI Z Y,GU J,YUN J,et al.Hypertensive cardiac remodeling effects of lignan extracts from Eucommia ulmoides Oliv.bark:A famous traditional Chinese medicine[J].The American Journal of Chinese Medicine,2003,41(4):801-815.
[7] 李琰,张靖,辛转霞,等.杜仲组培快繁的研究[J].西北农林科技大学学报(自然科学版),2004,23(6):79-82.
[8] 罗丽,赵德刚.杜仲带芽茎段的组织培养[J].江苏农业科学,2007(5):174-176.
[9] 唐亮,金晓玲.杜仲组织培养的研究进展[J].贵州农业科学,2010,38(3):15-18.
[10] 孟长军.组培实验室常见消毒方法优劣比较[J].中国园艺文摘,2012,28(8):177-178,168.
[11] 黄作喜,邱超,曾桢迦,等.植物组织培养中消毒剂的应用研究进展[J].内江师范学院学报,2013,28(6):26-30.
[12] 李素华,何松林.基本培养基和激素对常青白蜡腋芽诱导的影响[J].贵州农业科学,2009,37(12):175-176.
[13] 郑永春,迟丽华.笃斯越橘组培苗增殖扩繁培养基配方筛选试验[J].北方园艺,2013(9):138-139.
[14] 孙清荣,王金政,薛晓敏,等.‘金凯特’杏的组织培养与快繁[J].山东农业科学,2016,48(12):25-28.
[15] 德永军,吕涛,王一,等.文冠果茎段形成层不定芽诱导组织培养技术初探[J].内蒙古农业大学学报(自然科学版),2014,35(2):39-42.
[16] 张桂琴,徐祥龄,赵志学.文冠果嫩茎组织诱导植株移栽初获成功[J].林业科技通讯,1980(7):4-5.
[17] 张娜,张芸香,郭晋平.文冠果腋芽诱导关键影响因素与培养体系优化[J].山西农业大学学报(自然科学版),2014,34(1):53-58.
[18] 李琰,张存莉,严忠海,等.不同培养基对杜仲愈伤组织诱导和生长的影响[J].西北林学院学报,2003(3):37-39.
[19] 高国训.植物组织培养中的褐变问题[J].植物生理学通讯,1999(6):501-506.
[20] 黄烈健,王鸿.林木植物组织培养及存在问题的研究进展[J].林业科学研究,2016,29(3):464-471.
[21] 邢阿宝,崔海峰,俞晓平,等.光质及光周期对植物生长发育的影响[J].北方园艺,2018(3):163-172.
[22] SKOOG F,MILLER C O.Chemical regulation of growth and organ formation in plant tissues cultured in vitro[J].Symp Soc Exp Biol,1957,54:118-130.
[23] 谷文英,裴德清,朱登云,等.杜仲茎段培养的初步研究[J].莱阳农学院学报,1999(1):6-9.