白旗兜兰种子非共生萌发与试管苗快速繁育

对我国极小种群野生植物白旗兜兰的种子非共生萌发与试管苗快速繁育开展研究,对比了不同培养基、预处理、种子成熟度及光照对种子萌发的影响。白旗兜兰种子萌发的最适条件是授粉后270 d的种子,在1/4MS+10%椰汁培养基上,用1%Na OCl溶液处理种子40 min并在播种后的前4周进行黑暗培养。最适合原球茎转化幼苗的培养基是3.0 g·L-1花宝一号+1.0 mg·L-1NAA+0.5 g·L-1活性炭+10%香蕉匀浆,而最佳幼苗生根壮苗的培养基是3.0 g·L-1花宝一号+1.0 mg·L-1NAA+1.0 mg·L-16-BA+0.5 g·L-1活性炭+10%香蕉匀浆。     

多花兰种子无菌萌发及离体快繁研究

以多花兰种子为材料,研究了无机盐浓度、植物生长调节剂和光照条件对多花兰种子非共生萌发的影响,在此基础上,通过研究原球茎增殖和分化、芽苗壮苗和生根的培养基配方及培养条件,建立多花兰组培快繁技术体系.结果表明:多花兰种子萌发培养基为1/6 MS十NAA0.5 mg·L-1+6-BA 2.0 mg· L-1+马铃薯泥50g·L-1+AC 1.0g·L-1,光照度为1.25μmol·m-2·s-1,萌发率63.6％;原球茎增殖继代培养基为1/4 MS+6-BA2.0 mg·L-1+NAA 0.5 mg· L1+AC 1 g·L-1+PE 200 g·L-1,繁殖倍数6.5倍/60 d,芽分化率60.2％;再生芽分化培养基为1/4 MS十6-BA2.0mg·L-1+NAA 0.2 mg· L-1+AC 1 g·L-1+PE 200g· L-1,繁殖倍数4.0倍/60 d,芽分化率85.0％;芽苗壮苗和生根培养基为1/6 MS+6-BA 3.0 mg·L-1+NAA 1.0 mg·L-1+AC 1 g·L-1+蔗糖20g·L-1 +PE 200 g·L-1和1/4 MS+6-BA 2.0 mg·L-1+NAA1.2 mg·L-1+AC1 g·L-1十蔗糖20g·L-1+PE200g· L-1,生根率达100％,生根苗移栽成活率90％.此技术可用于多花兰种苗繁育和种质资源保护.     

濒危植物盘龙参种子的非共生萌发及种苗的快繁研究

以盘龙参种子为材料,筛选离体条件下适宜种子非共生萌发、种苗快繁的培养基。结果表明:盘龙参种子在无植物生长物质的培养基中能够萌发,但不能发育成苗;在含有1.0mg·L-1 KT、0.1mg·L-1 IAA和0.1mg·L-1 GA3的培养基中萌发,并能进一步发育形成种苗;在较高浓度生长素(1.2mg·L-1 NAA)的培养基中不能萌发。种苗在较高浓度细胞分裂素和较低浓度生长素配比的培养基中能够增殖,最高增殖系数可达到2.8,转入壮苗生长培养基中培养80d后可以移栽温室。最适增殖培养基为1/2MS+12.0mg·L-1 6-BA+0.1mg·L-1 NAA+10.0mg·L-1腺嘌呤;最适壮苗生根培养基为1/2MS+1.0mg·L-1 KT+0.1mg·L-1 IAA+10.0mg·L-1腺嘌呤。     

流苏树的组织培养和快速繁殖

以流苏树种胚为试材,研究不同种类及浓度的植物生长调节剂对其无菌体系建立,带顶、侧芽茎段伸长以及生根的影响。结果表明,种胚生根最适培养基为WPM+0.1 mg·L-1 NAA+3.0 mg·L-1 GA3+2.0 mg·L-1 6-BA,生根率为94.2%;带顶、侧芽茎段最适培养基为WPM+1.0 mg·L-1 GA3+1.0 mg·L-1 6-BA+0.5 mg·L-1 TDZ;生根最适培养基为WPM+0.1 mg·L-16-BA+0.5 mg·L-1 NAA+0.5 mg·L-1 IBA。     

爱沙木的组织培养和快速繁殖

1植物名称爱沙木(Eremophila bignoniiflora). 2材料类别无菌萌发的无根种子苗. 3培养条件种子萌发培养基:(1)MS 6-BA 1.0mg·L-1(单位下同) NAA 0.1.分化培养基:(2)MS 6-BA 2.0 NAA 0.1;(3)MS 6-BA 0.5 NAA0.1;(4)MS 6-BA 2.0 IBA 0.01;(5)MS 6-BA0.5 IBA 0.01.壮苗培养基:(6)MS.生根培养基:(7)1/2MS NAA 0.1 1%活性炭;(8)1/2MS NAA0.2 1%活性炭;(9)1/2MS IBA 0.01 1%活性炭;(10)1/2MS IBA 0.02 1%活性炭.以上除生根培养基加入20g·L-1蔗糖外均加入30 g·L-1蔗糖、7g·L-1琼脂,pH 5.6～5.8.光照12 h·d-1,光照度1 500～2000 lx,培养温度23～25℃.     

梳唇石斛成熟胚的离体培养和植株再生

1植物名称梳唇石斛(Dendrobium strongylanthum Rchb.f.). 2材料类别成熟种子. 3培养条件以MS和1/2MS为基本培养基.(1)种子萌发培养基:MS NAA 0.2 mg·L-1(单位下同) 6-BA 0.4 马铃薯汁200g·L-1;(2)原球茎诱导增殖培养基:1/2MS 6-BA 0.5;(3)原球茎分化培养基:1/2MS 6-BA 0.5 NAA 0.5 椰子汁200g·L-1 ;(4)壮苗及生根培养基:1/2MS NAA 0.5 香蕉泥100g·L-1 .上述培养基均附加20 g·L-1 蔗糖、8 g·L-1琼脂,pH 5.5～5.8.培养温度为(24±2)℃,光照时间12 h·d-1,光照度1 000～2000 lx.     

小白及的组织培养与快速繁殖

1植物名称小白及(Bletilla formosana). 2材料类别 种子. 3培养条件基本培养基为MS.(1)种子萌发培养基：MS KT 1.0～3.0mg.L-1(单位下同) NAA 1.0～3.0;(2)原球茎增殖培养基:1/2MS 6-BA 2.0;(3)原球茎分化培养基:1/2MS 6-BA 2.0 NAA 0.5;(4)生根培养基:1/2MS NAA 0.5.上述培养基均附加30g·L-1蔗糖、8 g·L-1琼脂,pH 5.8左右,培养温度为25℃左右,光照12 h·d-1,光照度2 0001x.     

火焰兰杂交种的胚培养和离体快繁

1植物名称火焰兰杂交种(Renanthera CoCCinea×R.imschootiana). 2材料类别种子. 3培养条件种子萌发培养基:(1)VW;(2)VW 椰子乳100mL·L-1;(3)KC;(4)KC 椰子乳100mL·L-1;(5)VW 椰子乳100mL·L-1 活性炭2 g·L-1(6)1/2MS 椰子乳100 mL·L-1.叶片离体培养基:(7)VW 2,4-D 1.0 mg·L-1(单位下同) 6-BA 2.5 NAA0.2;(8)VW 2,4-D 2.0 6-BA 5.0 NAA 0.5.原球茎继代增殖:(9)花宝1号1.5 g·L-1 花宝2号1.5g·L-1 椰子乳100 mL·L1 6-BA 1.0 NAA 1.0;(10)VW 椰子乳100mL·L-1 6-BA 1.0 NAA 1.0.生根壮苗培养基:(11)花宝1号3 g·L-1 蛋白胨2 g·L-1 活性炭2 g·L-1 NAA 0.5 6-BA 0.2;(12)花宝1号1 g·L-1 花宝2号1 g·L-1 蛋白胨2 g·L-1 活性炭2g·L-1 NAA 0.5 6-BA 0.2.以上培养基均加1.5%蔗糖、0.6%琼脂,pH 5.2～5.4,培养温度(25±2)℃,光照度1 500～2000 lx,光照时间12 h·d-1.     

短距槽舌兰的驯化栽培与快速繁殖

为促进短距槽舌兰的资源驯化栽培, 本文对其生物学特性、生长发育规律和组织培养快速繁殖技术进行了研究。对开花植株进行人工授粉, 取种子进行观察发现, 授粉后140 d 种子才达到成熟状态; 将其接种在无植物生长调节剂的RE培养基上, 萌发率可达90%以上。将萌发的种子转接在RE＋0.2 mg·L-1 NAA＋0.2 mg·L-1 6-BA＋5 g·L-1椰粉＋2 g·L-1水解酪蛋白的培养基上可诱导原球茎大量增殖。最适壮苗生根培养基为RE＋0.6 mg·L-1 NAA＋0.2 mg·L-1 6-BA＋80 g·L-1香蕉泥, 生根率100%。试管苗移栽到直径0.3-1.7 cm的小号细树皮盆栽中, 成活率在95%以上, 试管苗栽培6个月后可分化花芽, 这为短距槽舌兰的产业化生产奠定了基础。     

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%.     

印度梨形孢对铁皮石斛种子萌发和原球茎生长的影响

为探究印度梨形孢(Piriformospora indica)对铁皮石斛(Dendrobium officinale)种子萌发和原球茎生长的影响,在铁皮石斛种子离体培养和原球茎生长阶段分别接种印度梨形孢,对其形态发育特征和生理特性进行研究.结果表明,接种印度梨形孢的铁皮石斛种子的起始萌发时间提前,接种印度梨形孢的铁皮石...     

亚美万代兰与多花兰种子发芽过程的扫描电镜观察

兰种子发芽之后形成原球茎与根状茎,前者以亚美万代兰,后者以多花兰为材料进行扫描电镜观察。多花兰种子发芽初期,与亚美万代兰相似,但分化子叶的能力较差,在暗培养下,只形成鳞片状叶,紧贴在根状茎的分生组织上。根状茎只有转入光培养才有茎叶分化。兰种子发芽时的毛状物,在电子显微镜下观察,其形态与根毛较接近。     

同色兜兰的非共生萌发与快速繁殖研究

以授粉后不同发育时期的同色兜兰种子为材料,观察其形态特征和萌发过程,并探讨建立同色兜兰高效快繁体系的最佳条件。结果表明,种子发育中后期即授粉后210~240d为较适宜的采收期,授粉后210d的种子萌发率最高(达77.79%);1/4 MS和1/2MS为同色兜兰适宜的基本培养基,添加100mL/L椰乳或1g/L蛋白胨对种子萌发及原球茎生长和分化有明显的促进作用;添加1g/L活性炭对原球茎褐化有一定的抑制作用,但添加剂量不宜过大;添加香蕉汁和苹果汁对同色兜兰种子萌发和原球茎生长分化有抑制作用;暗处理对同色兜兰种子萌发无影响;分化后的原球茎在壮苗和生根培养基上培养120d即可得到4~5片叶、高3~5cm的同色兜兰健壮试管苗。     

金线兰种子萌发及其原球茎的增殖培养

研究了影响金线兰种子非共生萌发的因素,并应用正交试验研究基本培养基、6-BA、ZT、NAA四种因素对原球茎增殖的作用。结果表明:授粉类型对金线兰种子非共生萌发影响较大,异株异花、同株异花以及同株同花授粉所得的种子的萌发率分别为78.53%、69.62%、39.87%;蒴果成熟度以生长150d为宜,采收后萌发率可达78.59%;冷藏影响种子的活力,种子的萌发率随冷藏时间的延长而降低;使用次氯酸钠浸泡后的种子与对照相比,其萌发率无明显差异;NAA对原球茎增殖作用显著,适宜于原球茎增殖的培养基为1/2MS+ZT0.5mg/L+NAA1.0mg/L。     

Effects of mycorrhizal fungi on symbiotic seed germination of Pecteilis susannae (L.) Rafin (Orchidaceae), a terrestrial orchid in Thailand

Symbiotic seed germination of Pecteilis susannae (L.) Rafin was investigated using 11 fungal isolates recovered from roots of four Thai terrestrial orchids (P. susannae, Eulophia spectabilis, Paphiopedilum bellatulum and Spathoglottis affinis). Seed germination and protocorm development were evaluated up to 133 days after sowing. Protocorm development was most advanced, up to stage 5 (elongation of the first leaf), when seeds were cultured with 4 Epulorhiza isolates obtained from roots of P. susannae (CMU-Aug 028, 4.3%, CMU-Aug 007, 4.2%, and CMU-Aug 013, 2.2%) and E. spectabilis (CMU-STE 014, 3.9%). Moreover, stage 4 protocorm development (emergence of the first leaf) occurred with fungal isolates CMU-STE 011, 5.7%, (Epulorhiza sp.) and CMU-AU 212, 4.3%, (Tulasnella sp.) obtained from roots of E. spectabilis and S. affinis respectively. When seed was incubated without fungi (control), development was limited to stage 3 of protocorm development (appearance of promeristem). This is the first report of protocorm stage 5 development in P. susannae using compatible fungal symbionts. Optimization of seed germination and seedling fitness will assist the conservation and propagation of this orchid species and other terrestrial orchids in Thailand.     

In situ seed baiting to isolate germination-enhancing fungi for an epiphytic orchid,Dendrobium aphyllum (Orchidaceae)

Orchid conservation efforts, using seeds and species-specific fungi that support seed germination, require the isolation, identification, and germination enhancement testing of symbiotic fungi. However, few studies have focused on developing such techniques for the epiphytes that constitute the majority of orchids. In this study, conducted in Xishuangbanna Tropical Botanical Garden, Yunnan, China, we used seeds of Dendrobium aphyllum, a locally endangered and medicinally valuable epiphytic orchid, to attract germination promoting fungi. Of the two fungi isolated from seed baiting, Tulasnella spp. and Trichoderma spp., Tulasnella, enhanced seed germination by 13.6 %, protocorm formation by 85.7 %, and seedling development by 45.2 % (all P?Epulorhiza, another seed germination promoting fungi isolated from Cymbidium mannii, also enhanced seed germination (6.5 %; P?P?Trichoderma suppressed seed germination by 26.4 % (P?Tulasnella was the only treatment that produced seedlings. Light increased seed imbibition, protocorm formation, and two-leaved seed development of Tulasnella inoculated seeds (P?Tulasnella be introduced for facilitating D. aphyllum seed germination at the protocorm formation stage and that light be provided for increasing germination as well as further seedling development. Our findings suggest that in situ seed baiting can be used to isolate seed germination-enhancing fungi for the development of seedling production for conservation and reintroduction efforts of epiphytic orchids such as D. aphyllum.     

Morphogenesis of the protocorm ofCypripedium acaule (Orchidaceae)

The nature of the protocorm of theOrchidaceae has fascinated morphologists for more than a century. In the present study, the development of the protocorm was followed using in vitro germination of seeds on a culture medium containing sugar, but without a symbiont. Inside the seed, the embryo consists of about a hundred cells. In the embryo, cells are arranged along a longitudinal axis according to size; these cells contain protein and lipid reserve material. In the first stages of seedling development, the embryo is transformed into a protocorm and meristematic tissue becomes organized into a meristematic dome (promeristem) at the anterior pole. This meristematic dome will give rise to a scale and the apex of the seedling. At first, the apex and the scale leaf develop synchronously. The development of the root always follows that of the apex. The study of the development of the seed ofCypripedium acaule showed that the protocorm is a distinct morphological system with respect to the rest of the cormus. The protocorm may be interpreted as an extension of the proembryonic stage.     

硬叶兰种子的迁地共生萌发及有效共生真菌的分离和鉴定

兰科植物的种子原地和迁地共生萌发技术是近年发展起来的开展兰科植物种子和共生真菌研究的有效方法。该研究对兰属(Cymbidium)附生植物硬叶兰(C. mannii)开展了种子的迁地共生萌发研究, 试图获得其种子萌发的有效真菌。利用硬叶兰成年植株根部周围的树皮、苔藓、枯枝落叶、腐殖质等作为培养基质, 进行种子的共生培养。在培养133天后, 成功地获得了处于不同阶段的已萌发种子、原球茎和幼苗, 并从原球茎中分离得到一种瘤菌根菌属(Epulorhiza)真菌。用所分离到的FCb4菌株和一种从兜唇石斛(Dendrobium aphyllum)分离到的胶膜菌属(Tulasnella) FDaI7菌株和硬叶兰种子在燕麦琼脂培养基上进行共生萌发, 设置不接菌作为对照处理, 以检验FCb4菌株对硬叶兰种子萌发的有效性。经过58天的培养, 不接菌的对照处理中种子没有萌发, 接种FCb4和FDaI7菌株的处理都有很高的种子萌发率, 两种接菌处理在不同光照条件下的种子萌发率均无显著性差异。但暗培养条件下, 种子萌发形成原球茎后, 表现出生长停滞的趋势, 仅有很少的原球茎继续生长达到幼苗阶段, 说明原球茎发育后期与幼苗发育阶段需要光照。在光照条件下, 接种FCb4菌株处理中达到幼苗阶段种子的比例为(25.67 ± 9.27)%, 显著高于接种FDaI7菌株处理的(3.04 ± 2.27)% (W = 56, p = 0.026, Mann-Whitney U-test), 表明此研究中分离到的瘤菌根菌属真菌能有效地促使硬叶兰种子萌发并生长发育到幼苗阶段。     

In vitro propagation and mass scale multiplication of a critically endangered epiphytic orchid, Gastrochilus calceolaris (Buch.-Ham ex J.E.Sm.) D.Don. using immature seeds

In vitro asymbiotic seed germination potential of its immature seeds (36 weeks after pollination) of G. calceolaris was successfully tested on three different agar gelled nutrient media i.e. Murashige and Skoog (MS), Mitra et al. (M) and potato dextrose agar (PDA). Seeds germinated within 15.75+/-0.75 to 35.75+/-0.75 days in the three different media. The protocorms developed therefrom subsequently differentiated into first leaf and root primordia, and complete seedlings were obtained within 111.25+/-1.25 to 141.25+/-1.25 days on MS and M media. The protocorms, though failed to differentiate further on basal PDA medium, despite repeated subculturings, incorporation of peptone (P; 1 gl(-1)), yeast extract (YE; 2 gl(-1)) and coconut water (CW; 20%) in the medium proved beneficial in inducing differentiation, in these germinating entities. Additional use of growth additives (P/YE/CW), in general, favoured better germination, protocorm formation and seedling development. The optimal nutritional combination during seed germination, protocorm growth and multiplication and seedling development was found to be CW (10%) enriched MS medium.     

濒危莲瓣兰杂交育种及原生地种子萌发的研究

以莲瓣兰'大雪素'(Cymbidium tortisepalum 'Daxuesu')和'剑阳蝶'(Cymbidium tortisepalum 'Jianyangdie')为亲本,研究莲瓣兰种间杂交育种、杂交种子在原生地播种共生萌发及植株形态发生过程,为莲瓣兰杂交育种新品种选育及杂交种子原生地播种种苗快速繁殖奠定基础.结果表明,莲瓣兰种间杂交结实率在90.0%～93.3%之间,杂交亲和性较强;莲瓣兰杂交种子在原生地播种6～12月后,能被萌发菌侵染、种胚突破种皮形成类原球茎,种子在原生地播种容易萌发;类原球茎转绿成根状茎、根状茎伸长成丛生型根状茎、丛生型根状茎顶端分生组织分化出原球茎、原球茎分化出叶和根、根状茎逐渐退化完成植株再生过程极其缓慢,需要4～6年的时间.