Establishing Tissue Culture and Regeneration System of Scutellaria regeliana

doi:10.7525/j.issn.1673-5102.2020.01.008

Abstract

Abstract: We studied the effects of different disinfectant treatments and different plant growth hormone ratios on axillary bud induction, callus induction, cluster bud differentiation, proliferation, rooting and transplantation of Scutellaria regeliana stem segments with the stem segments of S.regeliana as explants. The best disinfection method was 0.1% HgCl₂ for 5 min, with the lowest pollution rate of 8.25%; the best medium for inducing axillary buds was MS+1 mg·L^-1 6-BA+1 mg·L^-1 NAA, with the induction rate of 73.66%; the best medium for inducing callus was MS+1 mg·L^-1 6-BA+1 mg·L^-1 2-4D, with the induction rate of 91.33%; and the best medium for inducing callus differentiation was MS+1 mg·L^-1 6-BA+0.5 mg·L^-1 NAA, with the differentiation rate of 44.71%. The optimum medium for bud multiplication was MS+1 mg·L^-1 6-BA+0.5 mg·L^-1 NAA, and the multiple of bud multiplication was 5.85; the optimum medium for rooting was 1/2MS+0.2 mg·L^-1 IBA, and the rooting rate could reach 74.07%; the volume ratio of vermiculite:perlite:pearlite:garden soil was 1:1:3, the highest survival rate of transplantation was 79.24%, and the plant grew vigorously. In this study, the regeneration system of S.regeliana was established to provide theoretical support in the development and application of S.regeliana wild resources on the basis of proper protection.

Key words: Scutellaria regeliana, adventitious bud, callus, regeneration system

CLC Number:

S567.239

ZHANG Jing, NIU Zhe, FAN Wei-Fang, GAO Peng-Fei, WU Jian-Hui. Establishing Tissue Culture and Regeneration System of Scutellaria regeliana[J]. Bulletin of Botanical Research, 2020, 40(1): 50-57.

References

[1] 贺美忠.黄芩组织培养与快速繁殖条件的研究[J].农业科技通讯,2016(4):79-80.He M Z.Study on tissue culture and rapid propagation conditions of Scutellaria baicalensis[J].Bulletin of Agricultural Science and Technology,2016(4):79-80.
[2] Yamamoto H,Chatani N,Watanabe K,et al.Effects of carbon sources on the growth and flavonoid formation of Scutellaria baicalensis stem callus cultures[J].Shoyakugaku Zasshi,1986,40(1):19-25.
[3] 高山林,陈柏君.黄芩组织培养快速繁殖技术的优化[J].中草药,2004,35(3):312-315.Gao S L,Chen B J.Optimization on cloning rapid-propagation technology by tissue culture in Scutellaria baicalensis[J].Chinese Traditional and Herbal Drugs,2004,35(3):312-315.
[4] 吴晓玲,胡海英,邓光存,等.黄芩愈伤组织诱导条件的研究[J].生物技术,2005,15(2):77-79.Wu X L,Hu H Y,Deng G C,et al.Study on inducing condition of callus of Scutellaria barcaleasis Georgi[J].Biotechnology,2005,15(2):77-79.
[5] 李富雄,张东向,李善文.黄芩同源四倍体的诱导及细胞形态学观察[J].北方园艺,2009(3):119-121.Li F X,Zhang D X,Li S W.The inducing of autotetraploid of Scutellaria baicalensis Georgi[J].Northern Horticulture,2009(3):119-121.
[6] 王淑华,李凤华,窦全丽.粘毛黄芩丛生芽的诱导及快速繁殖[J].安徽农业科学,2011,39(23):14022,14097.Wang S H,Li F H,Dou Q L.Induction and rapid propagation of Scutellaria viscidula shoots[J].Journal of Anhui Agricultural Sciences,2011,39(23):14022,14097.
[7] 薛欣,高山林,张汉超.黄芩无菌材料的建立及快繁技术的优化[J].药物生物技术,2011,18(4):332-335.Xue X,Gao S L,Zhang H C.Bacterium-free culture and rapid-propagation of Scutellaria baicalensis Georgi[J].Pharmaceutical Biotechnology,2011,18(4):332-335.
[8] 韩淑兰,王慧梅,张丹.降低黄芩再生苗玻璃化率及其生根的研究[J].植物研究,2016,36(1):90-96.Han S L,Wang H M,Zhang D.Decreasing the hyperhydricity and rooting of Scutellariae baicalensis Georgi[J].Bulletin of Botanical Research,2016,36(1):90-96.
[9] 刘占彬,袁庆华.多花黑麦草种子外植体组织培养灭菌方法研究[J].草地学报,2009,17(4):474-479.Liu Z B,Yuan Q H.Study on the sterilization methods in tissue culture of Italian ryegrass seed explant[J].Acta Agrestia Sinica,2009,17(4):474-479.
[10] 赵春莉,李金英.植物组织培养中污染原因及其控制方法[J].吉林农业科学,2010,35(6):11-13.Zhao C L,Li J Y.Reasons of contamination in plant tissue culture and controlling measures[J].Journal of Jilin Agricultural Sciences,2010,35(6):11-13.
[11] 汪腾越,周再知,裘珍飞,等.土沉香组织培养外植体消毒方法的研究[J].中南林业科技大学学报,2012,32(3):44-48.Wang T Y,Zhou Z Z,Qiu Z F,et al.Disinfection methods of explants of Aquilaria sinensis in tissue culture[J].Journal of Central South University of Forestry & Technology,2012,32(3):44-48.
[12] 孙健,王洪云,钮福祥,等.不同品种紫甘薯花青素含量及抗氧化活性差异[J].江苏农业科学,2013,41(12):323-324.Sun J,Wang H Y,Niu F X,et al.Anthocyanin content and antioxidant activity of different purple sweet potatoes[J].Jiangsu Agricultural Sciences,2013,41(12):323-324.
[13] Tsukuia A,Suzuki A,Komaki K,et al.Stability and composition ratio of anthocyanin pigments from Ipomoea batatas Poir.[J].Nippon Shokuhin Kagaku Kogaku Kaishi,1999,46(3):148-154.
[14] 吴翠平,沈学善,屈会娟,等.栽培因子调控马铃薯、甘薯等作物花青素合成研究进展[J].中国农学通报,2016,32(24):90-96.Wu C P,Shen X S,Qu H J,et al.Cultural factors regulating anthocyanin biosynthesis in potato,sweet potato and other crops[J].Chinese Agricultural Science Bulletin,2016,32(24):90-96.
[15] 吴春红,孔凡美,刘庆,等.氮肥对不同品种紫甘薯块根营养品质的影响[J].水土保持学报,2015,29(2):188-192.Wu C H,Kong F M,Liu Q,et al.Effects of different nitrogen levels on nutritional quality of different varieties purple sweetpotato storage roots[J].Journal of Soil and Water Conservation,2015,29(2):188-192.
[16] 胡佳卉,王小德.羊角槭愈伤组织诱导、增殖与分化[J].浙江农林大学学报,2018,35(5):975-980.Hu J H,Wang X D.Induction,proliferation,and differentiation of Acer yangjuechi callus[J].Journal of Zhejiang A&F University,2018,35(5):975-980.
[17] 何诗虹,王跃华,唐旭,等.羌活植物离体培养及植株再生研究[J].中药材,2017,40(7):1525-1528.He S H,Wang Y H,Tang X,et al.Study on in vitro culture and plant regeneration of Notopterygium incisum Ting ex H.T.Chang[J].Journal of Chinese Medicinal Materials,2017,40(7):1525-1528.
[18] 马均,马明东.曼地亚红豆杉的组织培养快繁技术[J].林业科学,2007,43(7):30-34.Ma J,Ma M D.Tissue culture technique of Taxus media ‘Hecksii’[J].Scientia Silvae Sinicae,2007,43(7):30-34.
[19] 张璐,潘远智,刘柿良,等.宜昌百合胚性愈伤组织诱导及植株再生体系的研究[J].植物研究,2019,39(3):338-346.Zhang L,Pan Y Z,Liu S L,et al.Embryonic callus induction and plant regeneration of Lilium leucanthum[J].Bulletin of Botanical Research,2019,39(3):338-346.
[20] 王盼盼.黑木相思不同优良无性系组培快繁技术比较研究[D].福州:福建农林大学,2011.Wang P P.A comparative study on the techniques of tissue culture and rapid propagation in different fine clones of Acacia melanoxylon[D].Fuzhou:Fujian Agriculture and Forestry University,2011.
[21] 唐佳佳,尚旭岚,洑香香.黑荆树不同外植体愈伤组织诱导与芽分化的研究[J].江苏林业科技,2014,41(2):6-10.Tang J J,Shang X L,Fu X X.Callus induction and differentiation with diverse explants of Acacia mearnsii[J].Journal of Jiangsu Forestry Science & Technology,2014,41(2):6-10.
[22] 李国福,陈士刚,秦彩云,等.垂枝桦组织培养技术研究[J].吉林林业科技,2010,39(3):1-3.Li G F,Chen S G,Qin C Y,et al.Study on tissue culture technology of Betula pendula[J].Journal of Jilin Forestry Science and Technology,2010,39(3):1-3.
[23] 李玲莉,郭素娟,李吉跃.北美柔枝松组织培养研究[J].北京林业大学学报,2010,32(3):209-212.Li L L,Guo S J,Li J Y.Tissue culture protocol of limber pine of North America[J].Journal of Beijing Forestry University,2010,32(3):209-212.
[24] 尹福良,刘丽彬,刘宝光.欧洲桤木组培快繁技术[J].林业实用技术,2014(1):28-29.Yin F L,Liu L B,Liu B G.Tissue culture and rapid propagation of Alnus cremastogyne[J].Practical Forestry Technology,2014(1):28-29.