铁皮石斛茎段快繁技术的研究

通过组织培养建立铁皮石斛快繁体系,为更好利用开发铁皮石斛提供原料。以铁皮石斛茎段为外植体,研究不同种类和浓度配比的激素和天然添加物对组培过程中不定芽的诱导和丛生芽的增殖的影响,筛选出各阶段最适培养基。结果表明,不定芽诱导最适激素配比为NAA 1.0 mg/L+BA 2.0 mg/L,丛生芽增殖最适培养基为NAA 0.5 mg/L+BA 1.0 mg/L+香蕉100 g/L+活性炭100 g/L。以茎段为途径建立铁皮石斛快繁体系高效稳定的获得组培苗是可行的。     

铁皮石斛同源四倍体工厂化快繁工艺研究

以实验室前期诱导的铁皮石斛四倍体为研究材料,利用气孔、叶绿体、染色体计数及流式细胞仪等方法,对铁皮石斛同源四倍体株系的纯合性进行鉴定,在此基础上利用茎段扩繁和原球茎诱导、增殖、分化两种方案比较增殖效率,并对原球茎的倍性稳定性进行再鉴定。结果表明：（1）实验室的‘花自 2’株系为纯合同源四倍体。（2）茎段扩繁最佳增殖培养基为MS＋2.0 mg/L 6 BA+0.5 mg/L NAA+15%香蕉汁,茎段诱导原球茎最佳培养基为MS+1.0 mg/L 6 BA+0.5 mg/L NAA+0.2 mg/L KT,诱导率达到76.66%;原球茎增殖最佳培养基为MS+1.0 mg/L 6 BA+0.5 mg/L NAA+15%马铃薯提取液,原球茎分化最佳培养基为1/2 MS+0.5 mg/L 6 BA+20%马铃薯提取液,原球茎分化率达到85.70%;生根最佳培养基为1/2 MS＋0.5 mg/L NAA＋15%香蕉汁,生根率达到92.77%。（3）原球茎根尖鉴定结果表明,诱导的倍性可以稳定遗传。（4）瓶苗移栽的最佳炼苗时间为5 d,移栽成活率达92.30%。该试验建立了铁皮石斛同源四倍体株系新的快繁技术体系,为四倍体试管苗的工厂化生产和推广奠定了技术基础。     

铁皮石斛组织培养与快速繁殖(简报)

以铁皮石斛种子为外植体进行组培快繁技术研究,筛选出适合各个阶段培养的培养基配方。种子萌发的培养基1/2MS+马铃薯汁15%;原球茎分化的培养基1/2MS+马铃薯汁20%+NAA 0.1 mg/L;壮苗培养基MS+香蕉汁10%+NAA 0.2 mg/L;生根培养1/2 MS+香蕉汁15%+NAA 0.5 mg/L。     

山嵛菜离体培养与快速繁殖研究

山嵛菜是一种重要的经济植物,用于医药、食品加工等领域。取山嵛菜带腋芽茎段作为外植体进行组织培养,利用长出的新芽直接诱导生成丛生芽。结果表明:MS+BA2.0mg/L+NAA0.05mg/L为最佳的丛生芽诱导和增殖培养基;生根较适宜的培养基为1/2MS+NAA0.05mg/L。     

Study on technology of aseptic sowing and rapid propagation of Demdrobium officinale

以铁皮石斛的蒴果为外植体,采用种子→原球茎→完整植株→移栽的途径快速成苗进行工厂化生产,对各阶段培养基进行筛选,以及其他一些影响因子进行比较研究.结果表明:人工授粉后生长60～180 d的铁皮石斛种子在离体条件下均能萌发,其中授粉150～180 d种子的萌发效果最好,萌发率为87.2%～94.4%,适宜的萌发培养基为MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+马铃薯汁200 g/L+AC 1.0 g/L;原球茎增殖的最佳培养基为MS+6-BA 1.5 mg/L+NAA 0.1 mg/L+香蕉汁100 g/L+AC 1.0 g/L,繁殖系数约为20倍/50 d;原球茎在MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+马铃薯汁200 g/L+AC 1.0 g/L培养基上进行分化培养,分化的同时还能进行一定的增殖;将已分化的芽苗转接到壮苗培养MS+6-BA 0.5 mg/L+NAA 0.2 mg/L+香蕉汁100 g/L+AC 1.0 g/L上培养1代后,转接到生根培养基1/2MS+NAA 0.8 mg/L+无机盐A 0.2～0.5 mg/L +香蕉汁100 g/L+AC 1.0 g/L上,培养50～70 d后,生根率100%,无机盐A可以有效地控制愈伤或原球茎的形成,明显提高生根苗的数量和质量.在桂林地区,生根苗以3～5月和9～10为最佳移栽期,以通过高温处理并堆沤腐熟的松树皮为基质,移栽成活率可达90%.     

铁皮石斛无菌播种产业化繁育技术研究

以铁皮石斛的蒴果为外植体,采用种子→原球茎→完整植株→移栽的途径快速成苗进行工厂化生产,对各阶段培养基进行筛选,以及其他一些影响因子进行比较研究。结果表明:人工授粉后生长60～180d的铁皮石斛种子在离体条件下均能萌发,其中授粉150～180d种子的萌发效果最好,萌发率为87.2%～94.4%,适宜的萌发培养基为MS+6-BA 1.0mg/L+NAA 0.1mg/L+马铃薯汁200g/L+AC 1.0g/L;原球茎增殖的最佳培养基为MS+6-BA 1.5mg/L+NAA 0.1mg/L+香蕉汁100g/L+AC 1.0g/L,繁殖系数约为20倍/50d;原球茎在MS+6-BA 1.0mg/L+NAA 0.1mg/L+马铃薯汁200g/L+AC 1.0g/L培养基上进行分化培养,分化的同时还能进行一定的增殖;将已分化的芽苗转接到壮苗培养MS+6-BA 0.5mg/L+NAA 0.2mg/L+香蕉汁100g/L+AC 1.0g/L上培养1代后,转接到生根培养基1/2MS+NAA 0.8mg/L+无机盐A 0.2～0.5mg/L+香蕉汁100g/L+AC 1.0g/L上,培养50～70d后,生根率100%,无机盐A可以有效地控制愈伤或原球茎的形成,明显提高生根苗的数量和质量。在桂林地区,生根苗以3～5月和9～10为最佳移栽期,以通过高温处理并堆沤腐熟的松树皮为基质,移栽成活率可达90%。     

铁皮石斛的组织培养(简报)

以铁皮石斛当年生茎段为材料,在MS＋6-BA 0.1 mg/L＋NAA 0.02 mg/L培养基上进行不定芽诱导培养;在MS＋6-BA 2.5 mg/L＋NAA 0.05 mg/L＋AC（活性炭）1g/L培养基上进行丛生芽诱导与增殖培养,50 d为一个继代周期,繁殖系数为5～8;在1/2 MS＋6-BA 0.5 mg/L＋NAA 0.05 mg/L＋椰子100 g/L＋AC 1g/L培养基上进行壮苗培养;在1/2 MS＋IBA 1.0 mg/L＋AC 1g/L培养基上进行生根培养;最后移栽到小颗粒化树皮中,成活率可达95%。     

不同植物生长调节剂对铁皮石斛(Dendrobium candidum)再生的影响

本实验以铁皮石斛无菌苗茎段为外植体,通过添加不同浓度的植物生长调节剂,诱导铁皮石斛愈伤组织形成与分化,建立铁皮石斛组织培养再生体系。结果表明,外植体接种5 d后,在改良MS培养基上添加2 mg/L PBU和0.05 mg/L IAA,可达到100%的愈伤诱导率。将愈伤组织接种在MS培养基上,添加0.1 mg/L NAA和0.5 mg/L 6-BA,不定芽诱导率为90.8%。适合铁皮石斛芽增殖培养基为:MS+7 g/L琼脂+30 g/L蔗糖+100 mg/L Vc+0.5 mg/L 6-BA+0.2 mg/L NAA。PBU浓度为0.5 mg/L,NAA浓度为0.05 mg/L时,芽苗生长情况良好,最适合用于不定芽伸长。铁皮石斛最适生根的培养基为:MS+7 g/L琼脂+30 g/L蔗糖+100 mg/L Vc+0.5 mg/L IBA,生根率可高达94.1%。本研究成功建立了铁皮石斛高效再生体系。     

铁皮石斛茎段诱导丛生芽的研究

以铁皮石斛Dendrobium candidum Wall.ex Lindl.的茎段作外植体,比较不同的培养基、激素等因素对茎段分化丛生芽的影响,以及不同浓度的香蕉汁和活性炭对试管苗生根的影响。结果表明：1／2MS较好,BA作用优于KT、ZT,最适浓度为2．0mg/L;丛生芽培养于1／2MS 10％香蕉汁 0．5％AC的培养基上,生根效果最好。     

太子参‘柘参1号’离体快繁试验

以太子参‘柘参1号’叶片、茎段为外植体进行离体快繁试验,结果表明：带腋芽的茎段能够诱导出丛生芽,且诱导率较高,最佳诱导培养基为MS + 6-BA 1.0 mg/L + NAA 0.2 mg/L +蔗糖30 g/L;增殖培养基为MS + 6-BA 1.0 mg/L + NAA 0.2 mg/L +蔗糖30 g/L;生根培养基为MS + NAA 0.2 mg/L +蔗糖30 g/L。     

红厚壳茎段丛生芽诱导与植株再生

红厚壳(Calophyllum inophyllum)为藤黄科红厚壳属多年生木本植物,有很高的药用价值。该研究以红厚壳带节茎段为外植体,探讨生长调节剂对腋芽萌发及丛生芽诱导、伸长和试管苗生根的影响。研究结果表明,外植体腋芽萌发和丛生芽诱导效果最好的培养基是MS+NAA1.0+TDZ0.5,在此条件下培养21天后,转入添加0.5 g·L–1活性炭且无生长调节剂的MS培养基,可有效促进不定芽的伸长。将带不定芽的外植体先在附加1.0 mg·L–1NAA的1/2MS培养基上进行生根诱导4周,之后转入附加1.0 g·L–1活性炭的无激素培养基进行根的伸长培养,这样的两步生根法能有效促进红厚壳生根。     

In vitro vegetative propagation of Chinese cabbage

Explants from cotyledons, axillary buds, inflorescence stems and flower buds of Brassica campestris ssp. pekinensis (Lour.) Olsson (Chinese cabbage, cv. Wongbok) were cultured on MS medium with growth regulators. Multiple shoots were obtained from cotyledons, axillary buds and flower buds but not from inflorescence stems. Propagation of shoots from cotyledons was more successful than from axillary buds and flower buds. The vegetative propagation rates varied amongst clones derived from cotyledons of the same cultivar and seed lot. The propagation rates of the cotyledon-derived material followed a normal distribution with an average propagation rate of 2.6 shoots per two weeks subculture when cultured on MS media plus 44.4 m benzyladenine (BA) and 14.8 m -indolebutyric acid (IBA). Shoots from three clones were cultured on MS medium with nine different concentrations of BA. The concentration of BA which promoted the highest rate of shoot propagation varied for the three clones and was in the range 44.4 to 177.6 m.     

澳洲青苹组织培养再生体系的研究

通过对澳洲青苹茎尖培养及植株再生体系中无菌培养物的建立、初代培养、继代培养、生根培养和移栽五个技术环节的研究,建立了澳洲青苹组织培养快速繁殖技术体系。结果表明：(1)BA0．3～0．6mg／L NAA0．1～0．3mg／L的MS培养基能促进澳洲青苹茎尖生长或大量侧芽的分化,不定芽生长健壮;(2)无根幼苗在含有IBA0．6～1．0mg／L NAA0．1mg／L的1／2MS培养基上暗培养一周后,再进行自然光培养,30d生根率可达到78%以上;(3)生根苗经50mg／L NAA浸渍1～2h后,移入蛭石：腐殖土：田园土=2：1：1的移栽基质中,保温保湿,成活率达80％以上;(4)进一步建立了自然光下生根、练苗一次完成的同步化体系,缩短了繁育时间,简化了步骤,降低了成本。此技术体系为澳洲青苹优良种苗快速繁殖提供了一条新途径。     

Effects of lead contamination on the clonal propagative ability of Phragmites australis (common reed) grown in wet and dry environments

Clonal propagation is important for the survival and maintenance of the common reed Phragmites australis. Pot culture experiments were conducted to investigate the effects of lead (Pb) concentration (0, 500, 1500, 3000, 4500 mg·kg^?1) and water stress on the clonal reproductive ability of this species. The Pb concentration found in plant organs, in decreasing order, was roots >shoots >rhizomes. There was a negative relationship between the growth of clonal propagative modules (excluding axillary shoot buds) and Pb concentrations, which caused a decrease in biomass, rhizome growth and number of axillary and apical rhizome buds. Daughter axillary shoots exhibited a tolerance strategy, with no significant change in their number; the axillary and apical rhizome buds, daughter apical rhizome shoots and rhizomes exhibited compensatory growth during the late stage of Pb (excluding 4500 mg·kg^?1) treatment in a wet environment. Pb applications above 500 mg·kg^?1 reduced these parameters significantly in the drought treatment, except for the number of axillary shoot buds, which did not change. Our results indicate that clonal propagative resistance to Pb contamination can occur via tolerance strategies, compensatory growth and a Pb allocation strategy, enabling these reeds to maintain population stability in wet environments. However, clonal modular growth and reproductive ability were inhibited significantly by the interaction between drought and Pb, which would cause a decline in P. australis populations in a dry environment. Lead concentrations of 4500 and 500 mg·kg^?1 in soils might meet or exceed the Pb tolerance threshold of clonally propagated reeds in wet and dry environments, respectively.     

柠檬马鞭草快速繁殖技术研究

选择柠檬马鞭草带腋芽的幼嫩茎段为外植体进行快速繁殖。结果表明,腋芽诱导的最适培养基是MS基本培养基附加6-BA0.5～1.0mg／L和IBA0．1～0．3mg／L．外植体腋芽能正常萌发生长,并迅速进入增殖状态,20d转接1次,增殖倍数存3倍以上,最适生根培养基是1／2MS基本培养基附加IBA0．5mg／L,诱导生根率达99％。将生根苗移入苗盆。30d的成活率在85％以上。     

野生毛葡萄远缘杂交后代离体培养研究

以MS、1/2MS、GS三种培养基为基本培养基,以及在1/2MS培养基上分别附加6-BA 1.0mg/L+IAA 0.1mg/L和6-BA 1.0mg/L+NAA 0.1mg/L两组植物生长调节剂,对野生毛葡萄及其远缘杂交种后代213、741、196、296、B₂等6个品种(种类)进行茎段腋芽萌发离体培养试验。结果表明:1/2MS培养基为基本培养基最适宜茎段腋芽的诱导萌发;NAA对毛葡萄及其杂交种后代的萌芽生长影响明显,若单独使用,对绝大多数品种的茎段腋芽萌发有明显的抑制作用,与IAA交替使用,则对茎段腋芽萌发有促进作用。     

Rapid in vitro propagation of Aloe barbadensis Mill

Axillary bud development and adventitious bud formation was obtained with decapitated shoot explants of Aloe barbadensis Mill. Maximal bud growth and rooting of shoots was obtained on a modified medium of Murashige and Skoog supplemented with 5 M IBA. More adventitious and axillary buds developed on nutrient media supplemented with IBA than with NAA. Axillary buds but not adventitious buds developed with IAA in the medium. Morphogenesis was inhibited by 2,4-D. Kinetin, benzyladenine and thidiazuron were toxic to the explants and did not stimulate the development of axillary of adventitious buds. The optimal temperature for bud growth and development was 25°C. Axillary bud growth and the formation of adventitious buds was slowed down at 10°C and totally inhibited by 30°C. The optimal sucrose concentration was 3% with the inhibition of bud growth and development by higher sucrose levels.     

In vitro micropropagation of the macarronesian evergreen treePersea indica (L.) K. Spreng

Summary Shoot propagation ofPersea indica (L.) K. Spreng was achieved using seedling axillary buds cultured on MS (Murashige and Skoog, 1962) medium with 1 mg/l (2.8 μM) N⁶-benzyladenine (BA). Forty percent of the obtained shoots did not elongate, but showed bud proliferation, which was maximal (three axillary buds per shoot) at the end of the seventh subculture. Sixty percent of the shoots elongated, did not show bud proliferation, and formed calluses at their base. Successful rooting (84.6%) was achieved dipping the base of each elongated shoot in 3 g/l (16.11 mM) indolebutyric acid (IBA) for 1–2 s, and transferring to half strength MS medium without growth regulators. These shoots presented an acclimatization success of 100%. Results suggest that micropropagated elongated shoots ofP. indica can be adequately used in reforestation programs.     

In vitro clonal propagation of Pisum sativum L.

A system of in vitro clonal propagation has been developed in Pisum sativum L. (cv. Bohatýr). A modified MS-medium supplemented with 20 M 6-benzylaminopurine (BAP) and 0.1 M -naphthaleneacetic acid (NAA) was used to induce multiple shoot formation from shoot apices, axillary buds of the first normal leaf, axillary buds of the first and second primary scales and axillary buds of cotyledons of 4 to 6 day old pea seedlings. Meristem explants maintained a high proliferation ability in each subculture in the course of 20 months of the culture. Regenerated shoots were rooted in the same basal medium containing 5 M NAA. Rooted plants were cultured in hydroponic pots filled with half-strength MS-medium to attain anthesis and seed maturity. The phenotypic uniformity of the regenerants was evaluated. Cytological investigation confirmed the diploid stage (2n=14) of regenerants and their progeny. Histological studies revealed that proliferating shoots originated from axillary and adventitious buds. In vitro propagation is discussed as related to pea breeding.