小花碱茅组织培养植株再生体系的建立

doi:10.11686/cyxb20140643

摘要/Abstract

摘要： 以小花碱茅成熟种子为外植体,研究了脱颖处理和不同激素配比对愈伤组织诱导和分化的影响,成功建立了其组织培养植株再生体系。结果表明,种子脱颖处理显著提高了种子发芽率,降低了污染率;诱导愈伤组织的最佳培养基为添加5.0 mg/L 2,4-D的MS培养基,诱导率可达55.2%,2周可见白色愈伤组织;最佳芽分化培养基为添加1.0 mg/L 6-BA和0.5 mg/L NAA的MS培养基,分化率可达31.7%,同时伴随根毛的发生,生根率达81.7%,诱导时间为2周;分化后的再生苗移植于1/2MS培养基上,100%生根,炼苗移栽后可全部成活。从小花碱茅种子诱导愈伤组织到植株再生共需16周左右。小花碱茅组织培养植株再生体系的建立为进一步探究小花碱茅耐盐碱分子机制奠定了基础。

Abstract: The effects of husk-off treatment and various hormone ratios on callus induction and differentiation were investigated using mature seeds of Puccinellia tenuiflora as explants. Finally, a plant regeneration system via tissue culture was established. The husk-off treatment increased seed germination rate and deceased contamination rate significantly. The optimal medium for callus induction was MS medium supplemented with 5.0 mg/L 2,4-D with the highest callus induction rate of 55.2%. The optimal medium for bud differentiation and rooting was MS medium supplemented with 1.0 mg/L 6-BA+0.5 mg/L NAA with the highest bud differentiation rate of 31.7%, the highest rooting rate of 81.7% within a period of two weeks. The rooting rate of regenerated plants was up to 100% on half strength MS medium and their transplant survival rate in soil was also up to 100%. The whole process of plant regeneration via tissue culture lasted for about 16 weeks totally. The establishment of plant regeneration system via tissue culture in P. tenuiflora has laid a foundation for further exploring the molecular mechanism of its salt tolerance.

中图分类号:

S812

王雪芳,王春梅,张金林,段丽婕,王锁民. 小花碱茅组织培养植株再生体系的建立[J]. 草业学报, 2014, 23(6): 355-360.

WANG Xue-fang,WANG Chun-mei,ZHANG Jin-lin,DUAN Li-jie,WANG Suo-min. Development of a plant regeneration system via tissue culture in Puccinellia tenuiflora[J]. Acta Prataculturae Sinica, 2014, 23(6): 355-360.

参考文献

Reference：
[1]Zhao X Y, Tong Z Y, Zhao B, et al.Cultivation of Puccinellia tehuifiora and its effects on improving alkaline spots of grasslands[J]. Pratacultural Science, 1992, 9(5): 61-62.
[2]Zhou B, Xu Z D. Reclamation and adaptability to saline - alkali meadow of Puccinellia Tenuiflora[J]. Jilin Normal University Journal(Natural Science Edition), 2005, 11(4): 30-31.
[3]Guo L Q, Chen J X, Cui J J, et al.Comparative studies of metabolic regulation of organic acids in Puccinellia tenuiflora under salt and alkali stresses[J]. Journal of Northeast Normal University(Natural Science Edition), 2009, 41(4): 123-128.
[4]Wei C X, Zhang J, Wang J J, et al. Observation on structural characters of vegetative organs of Puccinellia tenuiflora under salt stress[J]. Journal of Plant Resources and Environment, 2006, 15(1): 51-56.
[5]Wang S M, Zhu X Y, Wang Z R. A preliminary study on the effects osmotic adjustment on alkaligrass (Puccinellia tenuiflora) seedling growth under salt stess[J]. Acta Prataculturae Sinica, 1993, 2(3): 40-46.
[6]Wang S M, Zhu X Y, Shu X X. Studies on the characteristics of ion absorption and distribution in Puccinellia Tenuiflora[J]. Acta Prataculturae Sinica, 1994, 3(1): 39-43.
[7]Wang S M, Zhao G Q, Gao Y S, et al. Puccinellia tenuiflora exhibits stronger selectivity for K+ over Na+ than wheat[J]. Journal of Plant Nutrition, 2004, 27(10): 1841-1857.
[8]Guo Q, Wang P, Ma Q, et al. Selective transport capacity for K+ over Na+ is linked to the expression levels of PtSOS1 in halophyte Puccinellia tenuiflora[J]. Functional Plant Biology, 2014, 39: 1047-1057.
[9]Wang X, Yang R, Wang B C, et al. Functional characterization of a plasma membrane Na+/H+ antiporter from alkali grass (Puccinellia tenuiflora)[J]. Molecular Biology Reports, 2011, 38: 4813-4822.
[10]Ardie S W, Xie L, Takahashi R, et al. Cloning of a high-affinity K+ transporter gene PutHKT2;1 from Puccinellia tenuiflora and its functional comparison with OsHKT2;1 from rice in yeast and Arabidopsis[J]. Journal of Experimental Botany, 2009, 60: 3491-3502.
[11]Ardie S W, Liu S, Takano T. Expression of the AKT1-type K+ channel gene from Puccinellia tenuiflora, PutAKT1, enhances salt tolerance in Arabidopsis[J]. Plant Cell Reports, 2010, 29: 865-874.
[12]Zhao X, Li Y L,Zhang L X. Study on Zoysia seed germination characteristic and dormancy types[J]. Acta Botanica Boreali-Occidentalia Sinica, 2003, 23(11): 2003-2006.
[13]Ma C Y, Wang D, Ma J P, et al.Study on induction of callus from mature embryo and plant regeneration of Zoysia japonica[J]. Pratacultural Science, 2010, 27(9): 104-108.
[14]Li X L, Li K X, Yu X M. Establishment of Tissue Culture System for Puccinellia tenuiflora and Puccinellia chinampoensis[J]. Chinese Journal of Grassland, 2010, 11, 32(6): 90-93.
[15]Tang X Y, Yi Z L, Jiang J X, et al.Progress of plant hormone application in tissue culture of Ta1l Fescue[J].Hunan Agricultural Sciences, 2006, (3): 134-136.
[16]Ding L M, Long R J, Zhu T X.The Effect of 2,4-Dichlorophenoxyacetic Acid(2,4-D) and 6-Benzyladenine(6-BA) on Callus Inductionof Bluegrasses[J]. Grassland and Turf, 2003, 1: 34-37.
[17]Bregitzer P, Campbell R D, Wu Y. Plant regeneration from barley callus: effects of 2,4-dichlorphenoxy acetic acid and phenylacetic acid[J]. Plant Cell Tissue and Organ Culture, 1995, 43: 229-235.
[18]Sun Z H, Liu R T, Liang H M, et al.Progress of plant hormone application in tissue culture and plant regeneration of turfgrass[J]. Grassland and Turf, 2004, 2: 13-16.
[19]Qian H F, Xue Q Z. The Effect of Hormones on callus induction and plantlet regeneration of Tall Fescue (Festuca arundinacea)[J]. Chinese Journal of Grassland, 2002, 1(1): 46-60.
[20]Zhu X H, Sun J X, Liang H M, et al. Effect of low temperature and plant growth regulators on callus induction and regeneration of Zoysia japonica cv.Qingdao[J]. Pratacultural Science, 2009, 26(4): 121-126.
[21]Li J Q, Yun J F, Yun L, et al.Callus Induction of Immature Embryo of Psathyrostachys juncea[J]. Chinese Journal of Grassland, 2008, 30(2): 39-42.
[22]Xu D Y, He Y P. Effect of embryo treatment and different hormone concentration under dark culture on callus induction of Lolium perenne[J].Journal of Zhejiang Forestry Science and Technology, 2003, 23(2): 16-19.
[23]Yu G H, Ma H X, Yu J M, et al.Callus Induction and Plantlet Regeneration from Mature Seeds of Turf-type Tall Fescue[J].Jiangsu Journal of Agricultural Sciences, 2004, 20(1): 38-43.
参考文献：
[1]赵喜印, 佟振宇, 赵彬, 等. 星星草治理草原碱斑的效果与种植技术[J]. 草业科学, 1992, 9(5): 61-62.
[2]周波, 许志丹. 星星草对盐碱草地的适应性及改良作用的研究现状[J]. 吉林师范大学学报(自然科学版), 2005, 11(4): 30-31.
[3]郭立泉, 陈建欣, 崔景军, 等. 盐、碱胁迫下星星草有机酸代谢调节的比较研究[J]. 东北师范大学学报(自然科学版), 2009, 41(4): 123-128.
[4]韦存虚, 张军, 王建军, 等. 星星草营养器官适应盐胁迫的结构特征[J]. 植物资源与环境学报, 2006, 15(1): 51-56.
[5]王锁民, 朱兴运, 王增荣. 渗透调节在碱茅(Puccinellia tenuiflora)幼苗适应盐逆境中的作用初探[J]. 草业学报, 1993, 2(3): 40-46.
[6]王锁民, 朱兴运, 舒孝喜. 碱茅离子吸收与分配特性研究[J]. 草业学报, 1994, 3(1): 39-43.
[7]Wang S M, Zhao G Q, Gao Y S,et al. Puccinellia tenuiflora exhibits stronger selectivity for K+ over Na+ than wheat[J]. Journal of Plant Nutrition, 2004, 27(10): 1841-1857.
[8]Guo Q, Wang P, Ma Q,et al. Selective transport capacity for K+ over Na+ is linked to the expression levels of PtSOS1 in halophyte Puccinellia tenuiflora[J]. Functional Plant Biology, 2014, 39: 1047-1057.
[9]Wang X, Yang R, Wang B C,et al. Functional characterization of a plasma membrane Na+/H+ antiporter from alkali grass (Puccinellia tenuiflora)[J]. Molecular Biology Reports, 2011, 38: 4813-4822.
[10]Ardie S W, Xie L, Takahashi R,et al. Cloning of a high-affinity K+ transporter gene PutHKT2;1 from Puccinellia tenuiflora and its functional comparison with OsHKT2;1 from rice in yeast and Arabidopsis[J]. Journal of Experimental Botany, 2009, 60: 3491-3502.
[11]Ardie S W, Liu S, Takano T. Expression of the AKT1-type K+ channel gene from Puccinellia tenuiflora, PutAKT1, enhances salt tolerance in Arabidopsis[J]. Plant Cell Reports, 2010, 29: 865-874.
[12]赵昕, 李玉霖, 张立新. 两种结缕草种子休眠及萌发特性[J]. 西北植物学报, 2003, 23(11): 2003-2006.
[13]马彩云, 王栋, 马金萍, 等. 野生结缕草成熟胚愈伤组织诱导及再生体系的研究[J]. 草业科学, 2010, 27(9): 104-108.
[14]李晓玲, 李克秀, 于晓明. 星星草和朝鲜碱茅组织培养体系的建立[J]. 中国草地学报, 2010, 11, 32(6): 90-93.
[15]唐小艳, 易自力, 蒋建雄, 等. 植物激素在高羊茅组织培养中的应用与进展[J].湖南农业科学, 2006, (3): 134-136.
[16]丁路明, 龙瑞军, 朱铁霞. 2,4-D和6-BA对早熟禾愈伤组织诱导的影响[J]. 草原与草坪, 2003, 1: 34-37.
[17]Bregitzer P, Campbell R D, Wu Y. Plant regeneration from barley callus: effects of 2,4-dichlorphenoxy acetic acid and phenylacetic acid[J]. Plant Cell Tissue and Organ Culture, 1995, 43: 229-235.
[18]孙在红, 刘荣堂, 梁慧敏, 等. 植物激素在草坪草组织培养及植株再生中的应用与进展[J]. 草原与草坪, 2004, 2: 13-16.
[19]钱海丰, 薛庆中. 激素对高羊茅愈伤组织诱导及其分化的影响[J]. 中国草地, 2002, 1(1): 46-60.
[20]朱晓花, 孙吉雄, 梁慧敏, 等. 低温预处理与植物生长调节剂对结缕草愈伤组织诱导的影响[J]. 草业科学, 2009, 26(4): 121-126.
[21]李俊琴, 云锦凤, 云岚, 等. 新麦草幼胚愈伤组织诱导[J]. 中国草地学报, 2008, 30(2): 39-42.
[22]徐定炎, 何阳鹏. 种胚处理不同激素浓度在暗培养条件下对多年生黑麦草愈伤组织诱导的影响[J]. 浙江林业科技, 2003, 23(2): 16-19.
[23]余桂红, 马鸿翔, 余建明, 等. 草坪型高羊茅成熟种子胚性愈伤组织诱导及植株再生[J]. 江苏农业学报, 2004, 20(1): 38-43.
