水花生愈伤组织的诱导及根的分化
CALLUS INDUCTION AND ROOT DIFFERENTIATION FROM ALTENANTHERA PHILOXEROIDES
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摘要: 在1/2 MS培养基上培养出水花生的无菌苗,然后将水花生不同器官的外植体接种在不同培养基和添加不同种类激素及其浓度配比的培养基中,研究愈伤组织的诱导条件.结果表明:茎、叶为适宜组培的器官,苄基腺嘌呤(BA)、萘乙酸(NAA)、玉米素(ZT)、吲哚乙酸(IAA)的几种配比对水花生茎和叶的愈伤组织诱导均产生良好效应,NAA不变,随BA浓度上升,愈伤组织的诱导率上升.NAA/BA的比值越大越有利于根的分化.Abstract: With the development of modern industry and agriculture, a major problem faced by the modern world:environmental pollution. The lack of affordable,effective approaches to environmental remediation has created a major need for development of novel approaches. Plant have many endogenous genetic, biochemical,and physiological properties that make them ideal agents for soil and water remediation. Significant progress has been made in recent years in developing native or genetically modified plants for the remediation of environment contaminants. Improvement of plants by genetic engineering open up new possibilities for phytoremediation. Tissue culture is prerequisite to plant genetic engineering. Alternanthera philoxeroides may be useful in remediation of environmental pollutants by transgenic engineering. But there is no report on tissue culture of Alternanthera philoxeroides. Alternanthera philoxeroides is dicotyledon. It was collected from the suburb of YangZhou in JiangSu Province. In this paper, it were studied that effect of NaClO、AgNO3 and HgCl2 solution to sterilization of A. philoxeroides. Contamination rate were from 40% to 50% and germination rate were below 10% after stems with axillary bud of A. philoxeroides. were sterilized with 20% NaClO solution for 10—20 minutes. It was applicable to stems with axillary bud of A. philoxeroides to sterile with 1% AgNO3 solution for 20—25 minutes. contamination rate of A. philoxeroides were from 11.1% to 0 and germination rate were from 55 6% to 50% or to sterile with 0.1% HgCl2 solution for 3—5 minutes, contamination rate were from 25% to 0 and germination rate were from 58.3% to 44.4%.The results showed that the necessary time for killing bacteria contaminated in the tested material with 1% AgNO3 was 20—25 minutes of with 0.1% HgCl2 was 3—5 minutes. This study examined the effects of explants、media and exogenous phytohormone on tissue culture of Alternanthera philoxeroides . The stems,leaves and roots were used as the explants to study tissue cultue. Different basic media[MS、1/2MS、 MS(1/2)、B5],0—0.5mg/L of indoleacetic acid(LAA),0—0.2mg/L of naphthyl acetic acid(NAA),0.5—5.0mg/L of 6-benzylaminopurine(6-BA) and 0.5—4.0mg/L of Zeatin (ZT)as additional compositions were used for inducing, differentiating and rooting tests.Induced results showed that 1/2 MS was more suitable for calli to grow and divide than B5、MS(1/2)and MS. The stem and leaf of Alternanthera philoxeroides were suitable organs for tissue culture. Stem>leaf was observed on growth rate. The combinations of NAA and BA or IAA and ZT can induce calli of the stem and leaf of Alternanthera philoxeroides, and the rate of calli induction increases with the increase of BA concentration. The higher rate of NAA/BA,the more roots from the calli format. It was thought that the results of this study could provide an useful basis for genetic transformation of Alternanthera philoxeroides.
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Keywords:
- Alternanthera philoxeroides /
- Callus /
- Tissue culture /
- Root differentiation
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D,Peng X J.A research on the tolerance and purification ability of eight aquatic plants in heavy metal(Cu)contaminated sewage[J].China Environmental Science,1990,10(3):166-170[颜素珠,梁东,彭秀娟.8种水生植物对污水中重金属-铜的抗性及净化能力的探讨.中国环境科学,1990,10(3):166-170][3] Li X B,He G Y,Wu Z B,et al.Studies on some phytosynthetic parameters and chilling resistant enzymes in Eichhornia crassipes and Alternanthera philoxeroides[J].Acta Hydrobiologica Sinica,1995,19(4):333-337[李学宝,何光源,吴振斌,等.凤眼莲、水花生若干光合作用参数与酶类的研究.水生生物学报,1995,19(4):333-337][4] Xia H P,Q.L,Wang G.H.Kong.Phyto-toxicity of garbage leachates and effectiveness of plant purification for them[J].Acta Phytoecologica Sinica,1999,23(4):289-301[夏汉平,王庆礼,孔国辉.垃圾污水的植物毒性和植物净化效果之研究.植物生态学报,1999,23(4):289-301][5] Jia F Z,Xie P,Chen J L.Study of alternanthera philoxeroides for treating city sewage[J].Technology of Water Treatment,1998,24(5):308-310[贾凤芝,谢平,陈俊玲.水花生对城市污水净化能力的研究.水处理技术,1998,24(5):308-310][6] Xia H P.Uptake efficiency of Vetiveria zizanioides and Altehernanthera philoxeroides to N,P,Cl in garbage leachates[J].Acta phytoecologica Sinica,2000,24(5):613-616[夏汉平.香根草和水花生对垃圾污水中N、P、Cl的吸收效果.植物生态学报,2000,24(5):613-616][7] Liu J W,Lin F K,Wang Y.Study on purification ability of hydrophytes in Naphthalene contaminated water[J].Shanghai Environmental Sciences,2002,21(7):412-415[刘建武,林逢凯,王郁.水生植物净化萘污水净化能力研究.上海环境科学,2002,21(7):412-415][8] Guo S L,Huang C B,Bian Y.On absorption and accumulation of six heavy metal elements of six heavy metal elements of weeds in Jinhua subsurb-I Survey on content of six heavy metal elements in weeds and soil[J].Journal of Shanghai Jiaotong University(Agriculture Science),2002,20(1):22-29[郭水良,黄朝表,边媛.金华市郊杂草对土壤重金属元素的吸收与富集作用(Ⅰ)-6种重金属元素在杂草和土壤中的含量分析.上海交通大学学报(农业科学版),2002,20(1):22-29][9] Zhou C F,Wu G R,Shi G X,el al.The role of antioxidant systems in Cu2+ Stress resistance in Alternanthera philoxeroides[J].Acta Botanica Sinica,2001,43(4):389-394[周长芳,吴国荣,施国新,等.水花生抗氧化系统在低御Cu2+胁迫中的作用.植物学报,2001,43(4):389-394][10] Wang W J,Wang W F,Cao J J,et al.The induction of callus and plantlet regeneration from the leaf of wheat(Triticum aestivum L.)[J].Acta Bot.Boreal.-Occident.Sin,1998,18(3):401-405[王万军,王文芳,曹建军,等.小麦叶片愈伤组织及其再生植株的诱导.西北植物学报,1998,18(3):401-405][11] Wernicke W,Mickovits L.Development of gradient in wheat leaves response of leaf segments in different genotypes cultured in vitro[J].Plant physiology,1984,115:49-58[12] Rajarskarank MB Kei.Endogenous growth regulators in leaves and tissue culture of Pennisetum Purpureum scuum[J].Plant Physiology,1987,130:13-25[13] Wernicke W,Mickovits L.Rates of take up and metabolism of include-3-acetic and 2,4-dichlorophenoxyacetic acid by cultured leaf segment at different stages of devepment in wheat[J].Plant Physiology,1987,69:23-28[14] Pierik R.L.M.In vitro culture of Higher Plants[M].Dordrech:Martinus Nijhoff Publishers,1987,187-229[15] Gu R S,Jiang X N,Guo Z C.Advances in the studies on the mechanism of plant organogenesis in vitro[J].Chinese Bulletin of Botany,1999,16(3):238-244[谷瑞升,蒋湘宁,郭仲琛.植物离体培养中器官发生调控机制的研究进展.植物学通报,1999,16(3):238-244][16] Zhao J,Cheng J C.Effects of growth regulators on formation of callus,roots and bubs in Asparagus officinalis[J].Plant Physiology Communications,1991,27(4):253-255[赵洁,程井辰.生长调节物质对石刁柏愈伤组织诱导及根芽分化的影响.植物生理学通讯,1991,27(4):253-255][17] Xu Z H,Wang X,Liu G Y.Studies on organogenesis in the culture of the leaf explant of Nicotiana tabacumL[J].Acta Phytophysiologia Sinica,1978,4(2):177-182[许智宏,王雄,刘桂云.烟草叶组织培养中器官形成的研究.植物生理学报,1978,4(2):177-182]
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