影响喜树组织培养苗离体生根的因素

为了建立有效的喜树(Camptotheca acuminata)组培苗生根系统,提高其移栽成活率及适应性,用不同生长素种类及浓度、不同蔗糖浓度及不同培养基对喜树组培苗不定根形成影响以及移栽初期根系发育状况进行了研究.结果发现:1)生长素种类和浓度明显影响喜树组培苗不定根形成,在含有IBA0.5 mg·L-1培养基中取得了最佳生根效果,生根率达到了98%,外植体平均生根数为5.9条/株;2)不同浓度蔗糖对喜树组培苗生根也有一定影响,在10～30 g·L-1范围内,随着蔗糖浓度增加,生根百分率和生根数量都有增加,蔗糖浓度达到30 g·L-1时,生根百分率为95%,外植体平均生根数为5.4条/株;蔗糖浓度在40 g·L-1时,表现出对生根抑制作用;3)在基本培养基对喜树组培苗生根影响研究中发现,MS培养基对根形成表现出一定抑制作用;1/2MS和WPM培养基均适合喜树组培苗生根;4)根系发育正常的喜树组培苗移栽后成活率可达96%,但组培苗根系根毛系统发育较差.组培苗单位叶面积根尖数量显著低于对照实生苗,而且此参数与叶片气孔导度呈显著正相关.这种较差根系发育导致叶片气孔导度过低可能是组培苗叶片光合能力较低的重要原因.     

紫花亚菊（Ajania purpurea）的组织培养与快速繁殖

以紫花亚菊茎段为外植体对其进行组织培养,MS为基本培养基,设置不同激素浓度配比,对试验结果进行观察分析,筛选出适合的配方：启动培养基为MS＋6-BA0.5mg·L-1＋NAA0.01mg·L-1;继代培养基MS＋6-BA0.3mg·L-1＋NAA0.05mg·L-1,组培苗分化率高;不定根最适诱导培养基为：1/2MS＋IBA0.15mg·L-1,生根率达90%以上,组培苗移栽成活率达95%。     

樟叶越桔组培苗生根和移栽技术研究

为解决樟叶越桔(Vaccinium dunalianum)组培苗生根质量不佳、移栽成活率低的问题,该研究以樟叶越桔继代苗为材料,采用单因子试验从激素类型及浓度、培养基类型和蔗糖质量浓度对其生根的适宜条件进行筛选,进一步研究了不同基质配比对樟叶越桔移栽苗存活率的影响。结果表明:激素类型和浓度、培养基类型对樟叶越桔生根率的影响最大,其次为蔗糖质量浓度;最适合樟叶越桔生根的激素及浓度为IBA2.0 mg·L~(-1)、基本培养基类型为1/4MS、蔗糖质量浓度为15 g·L~(-1),樟叶越桔组培苗最佳生根培养基为1/4MS+IBA 2.0 mg·L~(-1)+活性炭0.1 g·L~(-1)+蔗糖15 g·L~(-1),生根率达100%,平均生根数为每株7.67条;根系呈辐射状、基部无愈伤组织,组培苗生长健壮、叶色浓绿;樟叶越桔组培苗移栽时以全腐殖土基质为佳,成活率为83.7%,植株叶片舒展,生长状况良好。该研究建立的优化体系有效地提高了樟叶越桔组培生根苗的生根率和生根质量,解决了后期移栽成活困难的问题,为优良的樟叶越桔植株规模化生产提供了科学依据和技术支持。     

万寿菊杂交亲本的离体培养

以万寿菊‘钻石’雄性不育株的幼嫩叶片和子房为外植体进行离体快繁,结果表明:培养基MS+0.8 mg·L-1 IAA+0.6 mg·L-1 6-BA+30 g·L-1蔗糖既有利于叶片愈伤组织的诱导也有利于不定芽的分化,愈伤组织诱导率达97.9％,不定芽诱导率达45.8％;培养基MS+0.5 mg·L12,4-D+0.5 mg·L-16-BA+ 30 g·L-1蔗糖为诱导子房愈伤组织的最佳培养基,出愈率为77.8％;培养基MS+0.05 mg·L -1 NAA+0.5 mg·L-1 6-BA+ 30g·L-1蔗糖为诱导子房愈伤组织不定芽分化的最佳培养基;将不定芽接至MS培养基上,7d后即可生出不定根,生根率可达98％,移栽成活率达90％以上.     

马来沉香组织培养技术研究

以马来沉香茎段为外植体,分别对外植体的消毒、启动培养、增殖培养、壮苗培养、生根培养、炼苗移栽环节进行研究,着重探索马来沉香组织培养技术各个环节的最佳培养基配方,为马来沉香的工厂化育苗提供技术指导。结果表明:马来沉香最佳消毒方法是用0.1%升汞消毒4～5min;启动率最高的培养基配方是1/2MS+6-BA 0.2mg·L-1+NAA 0.1mg·L-1+蔗糖30g·L-1+琼脂5.8g·L-1,启动率达70.5%;增殖系数最高的培养基是1/2MS+0.1mg·L-16-BA+25g·L-1蔗糖+5.8g·L-1琼脂,增殖系数达2.9;最佳壮苗培养基是1/2MS+30g·L-1蔗糖+5.8g·L-1琼脂;最佳生根培养基为1/2MS+NAA 5.0mg·L-1+20g·L-1糖+6g·L-1琼脂,培养2d后移入1/2MS培养基继续培养,生根率为83%;马来沉香移栽较难成活,在泥炭土∶黄泥土(2∶1)的基质上成活率最高,移栽成活率65%。     

单头亚菊的组织培养与快速繁殖

以单头亚菊茎段为外植体对其进行组织培养,MS为基本培养基,设置不同激素浓度配比。对实验结果进行观察分析,筛选出合适的配方。启动培养基为Ms＋0．5mg·L-16-BA＋0．01mg·L-1NAA。继代培养基MS＋O．75mg·L-1。6-BA＋0．01mg·L。NAA,可获得较高的增殖率。不定根最适诱导培养基为1／2MS＋O．15mg·L—IBA,生根率达87％以上,组培苗移栽成活率达98％。     

长筒石蒜鳞片诱导和植株再生

采用长筒石蒜带基盘鳞片为外植体,以MS为基本培养基,附加不同种类和浓度的植物生长调节物质诱导小鳞茎,在NAA 0.1 mg·L-1 6-BA 5.0 mg·L-1及ZT 0.5 mg·L-1 6-BA 5.0 mg·L-1的培养基上,小鳞茎(芽)增殖率可达480%;以蔗糖浓度为6%的MS培养基上的小鳞茎生长量最高;小鳞茎在MS培养基上生根率可达100%;移栽成活率为90%左右.     

不同因素对小黄李试管苗增殖与生根的影响

以‘小黄李’砧木的单芽茎段为外植体进行离体培养,研究不同培养基、植物生长调节剂的种类及其浓度、pH值对‘小黄李’增殖与生根的影响。结果表明:较合适的增殖培养基为F14 0.8mg·L-16-BA 0.2mg·L-1NAA 30g·L-1蔗糖 6g·L-1琼脂,pH值5.4~5.8;较合适的生根培养基为改良MS 0.1mg·L-1NAA 0.2mg·L-1IBA 30g·L-1蔗糖 6g·L-1琼脂,生根率达100%,单株平均根数3~4根;以蛭石∶珍珠岩∶泥炭(1∶1∶1)移栽生根苗,成活率达80%以上,实验结果为工厂化育苗生产奠定了基础。     

广西绞股蓝的组织培养和快速繁殖

广西绞股蓝是极具开发潜力的绞股蓝属植物。以其茎尖和叶片为外植体进行的组织培养试验表明,茎尖为最适的外植体,愈伤组织诱导和不定芽增殖的最适培养基为MS+6-BA 1.0mg.L-1+NAA 0.02mg.L-1,在此培养基中,茎尖和叶片的愈伤组织诱导率平均达95%,丛芽增殖系数平均达14。广西绞股蓝最适的生根培养基为MS+NAA 0.20mg.L-1,生根率92.50%。炼苗后组培苗的移栽成活率达95.5%。     

美洲黑杨(中嘉8号)离体叶片再生研究

用中嘉8号杨[Populus deltoids(I-63×I-69)]试管苗叶片作外植体,在添加不同生长素和细胞分裂素浓度配比的1/2(N)MS培养基上,筛选出叶片直接分化不定芽的最适培养基1/2(N)MS 0.2 mg/L BA 0.02 mg/LNAA 25 g/L蔗糖 7 g/L琼脂,不定芽分化率为85%。同时发现,一定浓度的KT对生根具有促进作用,最适生根培养基为MS 0.08 mg/L KT 0.02 mg/L NAA 25 g/L蔗糖 8 g/L琼脂,生根率为82.2%。     

海滨锦葵胚轴愈伤组织诱导及植株再生

以海滨锦葵(Kosteletzkya virginica)胚轴为外植体, 在9种不同激素配比的培养基上进行愈伤组织诱导、继代培养、不定芽分化及生根培养, 确定了植株再生的最适培养条件: (1)愈伤组织诱导最适培养基为MS + IAA 1.0 mg.L-1 + KT 0.3 mg.L-1 + sucrose 30 g.L-1 + agar 8 g.L-1, 愈伤组织诱导率为93.94%; (2)不定芽诱导最适培养基为MS + IAA 0.1 mg.L-1 + ZT 0.5 mg.L-1 + sucrose 30 g.L-1 + agar 8 g.L-1, 不定芽诱导率为65.83%; (3)生根最适培养基为MS + sucrose 30 g.L-1 +agar 8 g.L-1, 生根率为96.67%。炼苗移栽后, 成活率可达85%。     

Regeneration of niger (Guizotia abyssinica Cass.) CV Sahyadri from seedling explants

Root, hypocotyl and cotyledonary explants of niger (Guizotia abyssinica Cass) CV. Sahyadri were aseptically cultured on Murashige and Skoog's basal medium (MS) containing BAP and kinetin. Multiple shoot regeneration was induced from hypocotyl and cotyledonary explants while root explants produced only callus on MS medium supplemented with BAP. BAP (1 mg l^-1) was optimum for shoot regeneration. Regenerated shoots were transferred to MS medium without auxins, with auxins and with increasing concentrations of sucrose for rooting. Complete plantlets were obtained in all cases; however, 0.5 mg l^-1 NAA was the best for induction of roots. Ninety-seven per cent of the plantlets survived and completed their life cycle when transferred to natural conditions.Abbreviations BAP 6-benzylamino purine - NAA -naphthaleneacetic acid - IAA indole-3-acetic acid - IBA indole-3-butyric acid     

In vitro sucrose concentration affects growth and acclimatization of <Emphasis Type="Italic">Alocasia amazonica</Emphasis> plantlets

Plantlets of Alocasia amazonica were regenerated on the MS medium supplemented with different concentrations (0–9%) of sucrose. An absence of sucrose in the growth medium induced generation of leaves, however, it decreased multiplication. On contrary, sucrose supply of 6% or 9% enhanced multiplication but hampered photoautotrophic growth (generation of leaves). Increasing sucrose supply also increased sugars and starch content and number of stomata and decreased water potential and size of stomata during in vitro growth period. During ex vitro acclimatization, shoot length, root length, leaf number and root number of Alocasia plantlets grown with 3% sucrose, were found to be better among the other studied sucrose concentrations. Under ex vitro acclimatization, number of stomata, contents of various carbohydrates in the leaves were increased but size of stomata decreased with increasing sucrose supply during in vitro growth period. Moreover, water potential of leaves of plantlets, which have been grown with a sucrose concentration other than 3%, was decreased. During in vitro growth, net CO₂ assimilation rate (P_N), transpiration (E), stomatal conductance (g_s) and variable fluorescence to maximum fluorescence ratio (Fv/Fm) were unaffected, however, during acclimatization these were changed and maximum P_N, E, and g_s were observed in the plantlets micropropagated with 3% sucrose. Fv/Fm was decreased severely in the plantlets micropropagated with 6% sucrose during acclimatization. Thus a sucrose concentration of 3% in the medium is appeared to be better among studied concentrations for both in vitro growth and ex vitro acclimatization of A. amazonica plantlets.     

珍稀药用和观赏植物地涌金莲的组织培养和快速繁殖

以地涌金莲(Musella lasiocarpa)吸芽为外植体建立了有效的快繁体系。外植体经灭菌处理后在MS 6-BA4.0 mg L-1 NAA 0.2 mg L-1 维生素C 150 mg L-1 10%椰子乳 3%蔗糖培养基上能进行不定芽的诱导和增殖,培养60 d后每个芽平均能产生4.10个不定芽。第6代增殖后,丛生芽的增殖系数可达4.23。生根培养基以1/2MS NAA1.0 mg L-1 AC 50 mg L-1的效果较好。以沙:泥炭土:珍珠岩=1:1:1为基质移栽试管苗,成活率达到93.5%以上。经过12个月的组织培养已生产10 000多株试管苗。     

米老排的组织培养和快速繁殖

以米老排人工林成年优树当年生枝条茎段为外植体,建立了“以芽繁芽”的组织培养快速繁殖体系。丛芽诱导培养最快的无性系,经过3个月的培养,一个外植体可获得17．2个芽。在添加6-BA1．0mg·L-1的Ms培养基上增殖率最高,月增殖率为2．43。促进苗高生长的最有效培养基是Ms＋6-BA0．4mg·L-1＋GA30．4mg·L-1。生根培养基为1／2MS＋IBA0．4nag·L-1＋O．1g．L-1活性炭,生根率达81．8％以上,每株苗生根7．8条,平均苗高为1．2cm。经生根培养20d和目光温室炼苗15d后,试管苗移植入黄泥和泥炭土（4：1,刃功混合基质中,成活率达85％以上。     

Plant regeneration from leaf explants of Aloe barbadensis Mill. and genetic fidelity assessment through DNA markers

An efficient plant regeneration protocol was developed from leaf explants of Aloe barbadensis Mill on Murashige and Skoog’s (MS) medium supplemented with 2.0 mg/l 6-benzyladenine (BA) or Kinetin (Kn), 0.25–0.5 mg/l NAA (1-napthalene acetic acid) and 3 % (w/v) sucrose within 4 weeks of culture. The maximum number of shoot buds were obtained on MS medium supplemented with 2.0 mg/l BA, 0.5 mg/l NAA, 40 mg/l Ads (adenine sulphate) within 4–6 weeks of subculture. Inclusion of 0.25–0.50 mg/l gibberellic acid into the medium, the shoot buds became elongated. Repeated subculture on regeneration medium induces higher rate of shoot regeneration. The root induction from excised microshoots was achieved on half-strength MS medium supplemented with 0.25–1.0 mg/l NAA or indole-3-butyric acid (IBA) and 2 % (w/v) sucrose. Maximum percentage of rooting was achieved on medium having 0.5 mg/l NAA with 3 % (w/v) sucrose. About 80 % of in vitro raised plantlets were hardened in the greenhouse and successfully established in the soil. Both Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeat (ISSR) markers were used to detect the variability among the regenerated plants developed in vitro. The results showed that there was no polymorphism among the regenerated plantlets. This study will help for propagation of quality planting material of Aloe barbadensis for commercialization.     

In vitro propagation of an endemic and endangered medicinal plant <Emphasis Type="Italic">Notopterygium incisum</Emphasis>

An efficient system in vitro propagation for Notopterygium incisum Ting ex H. T. Chang, an endemic and endangered medicinal plant, was established to address increased demand and germplasm conservation goals. Optimum response in callus induction (CI) was observed on Murashige and Skoog (MS) medium supplemented with 1.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.2 mg/l 6-benzylaminopurine (BAP), which the induction rate and growth of callus were 84.44% and 0.67 g respectively. The highest shoot regeneration frequency (76.97%) and maximum number of shoots (3.6 shoots per callus) were achieved on MS medium with 1.5 mg/l BAP and 0.2 mg/l naphthalene acetic acid (NAA). Half-strength MS medium supplemented with 0.6 mg/l indole-3-butyric acid (IBA) was determined to be the best rooting medium, resulting in the maximum number of roots (18.6 roots per shoot) and the highest rooting frequency (92.28%). An approximate 83.8% survival rate among the regenerated plantlets was recorded after they were transplanted in the field at an altitude of 3200 m. An HPLC analysis showed that the content of two main chemical constituents, notopterol and isoimperatorin, in the rhizomes of 3-year-old regenerated plantlets was higher (3.84 mg/g and 4.05 mg/g, respectively) than that in commercially marketed crude drugs. This first report of complete regeneration in vitro could provide an alternative method for the rapid, large-scale production and conservation of this valuable, rare, and endangered medicinal plant.     

A rooting procedure for lentil (Lens culinaris Medik.) and other hypogeous legumes (pea,chickpea and Lathyrus) based on explant polarity

The present study assessed the rooting response of lentil nodal segments in relation to explant polarity, hormone, salt and carbohydrate concentrations of the medium. Nodal segments of lentil with an axillary bud cultured in an inverted orientation (apical end in medium) showed higher rooting frequencies than explants cultured in a normal orientation (basal end in medium). The highest rooting percentage (95.35%) and average number of shoots regenerated per explant (2.4) were obtained from explants placed in an inverted orientation on Murashige and Skoog (MS) medium salts with 3% sucrose, supplemented with 5 microM indole acetic acid (IAA) and 1 microM kinetin (KN). Reducing or increasing phytohormone concentration did not alter significantly root regeneration of inverted explants. Sucrose at 3% allowed higher root regeneration frequencies compared to 1.5% sucrose. MS full concentration permitted regeneration of longer shoots with more nodes per regenerated shoot, compared to MS half-strength, which regenerated more shoots of shorter length and with less nodes. Inverted nodal segments of other hypogeous legumes (pea, chickpea and Lathyrus) also exhibited higher rooting frequencies than explants cultured in a normal orientation on MS medium with 3% sucrose and supplemented with 5 microM IAA and 1 microM KN. The most novel application of this study is the culture of nodal segments of hypogeous legumes in an inverted orientation. This procedure is a considerable improvement over other published procedures concerning in vitro rooting of lentil, pea, chickpea and Lathyrus.     

铁核桃离体培养与快速繁殖

以优良单株‘纳雍-1’的单芽茎段为外植体,建立了铁核桃（Juglanssigillata）离体培养与快速繁殖的体系。结果表明,附加6-BA1．0mg·L-1 ＋活·IgK（AC）3．0g·L-1的DKw培养基适宜铁核桃腋芽诱导;适宜铁核桃芽增殖的培养基为DKW＋6-BA1．0mg·L-1 ＋IBA0．02mg·L-1,40d后增殖系数可达7．33;试管苗的茎尖和茎段均可用于增殖培养;一步生根法（低浓度的生长素IBA持续诱导）不利于铁核桃试管苗嫩茎生根;采用二步生根法,生根率最高可达71．73％,其中,不同IBA浓度、暗培养时间、蔗糖浓度和AC含量对试管苗嫩茎生根影响显著,铁核桃试管苗在附]sulBA5,0mg·L-1的1／4DKW培养基中暗培养12d,再转移到不含IBA的1／4DKW培养基（附加AC 3g-L-1和蔗糖20g·L-1）中生根效果最好;生根试管苗采用珍珠岩和营养土两步炼苗,60d后成活率达到87．50％。