扁蓿豆体细胞胚的诱导和植株再生

扁蓿豆实生苗的根、下胚轴、子叶、叶片和叶柄外植体,在含２,４—Ｄ２—０．２５ｍｇＬ－１与ＫＴ０．２５－２ｍｇＬ－１及２,４—Ｄ０．５ｍｇＬ－１与ＺＴ０．５ｍｇＬ－１或ＢＡＰ０．５ｍｇＬ－１与ＮＡＡ０．０５ｍｇＬ－１的ＭＳ琼脂培养基上均可产生愈伤组织．愈伤组织在含２,４—Ｄ０．５—０．１ｍｇＬ－１与ＫＴ０．５—０．１ｍｇＬ－１或ＢＡＰ０．２５＋ＮＡＡ０．０５ｍｇＬ－１的ＭＳ培养基上可诱导分化出体细胞胚．体细胞胚在无激素的培养基上发育成完整植株．用海藻酸钠包襄体细胞胚制成人工种子,其发芽率和植株转换率分别为９５％和５３％．     

AgNO3对大白菜子叶芽再生的促进作用

以大白菜“０２号杂交早皇白”无菌苗的子叶为材料,用附加ＢＡ２．０ｍｇＬ＾－１、ＮＡＡ０．１～１．０ｍｇＬ％－１的ＭＳ培养基培养,能直接放导分化 出芽,最适激素比例为ＢＡ２．０ｍｇＬ＾－１、ＮＡ０．５ｍｇＬ＾－１,芽的分化率为３１．６％,在上述培养基里活加２ｍｇＬ＾－１的ＡｇＮＯ３能使芽的再生频率提高至８６．５％。     

可乐茄叶肉原生质体培养再生植株

本文报道茄属果树可乐茄（ＳｏｌａｎｕｍｑｕｉｔｏｅｎｓｅＬａｍ．）叶肉原生质体的分离、培养及植株再生。幼嫩叶片原生质体经酶游离、纯化后,以１×１０４个／ｍｌ密度培养于稍加改良Ｋ８ｐ（附加２,４－Ｄ０．５ｍｇＬ（－１）、ＮＡＡ１．０ｍｇＬ（－１）和ＢＡ０．５ｍｇＬ（－１））的培养基中,三天后开始分裂,一周分裂３—４次。一个月形成小细胞团,植板率为０．１—０．２％,小细胞团转培养于ＭＳ＋２,４－Ｄ０．５ｍｇＬ（－１）上增殖后进行分化。原生质体来源愈伤组织在ＩＡＡ（０．１—１．０ｍｇＬ（－１））与ＢＡ或ＺＴ组合的培养基中能诱导器官发生,芽分化率最高可达４２．９％;但ＩＡＡ、ＢＡ、ＺＴ三者一起使用未见任何器官分化。小芽在ＭＳ＋ＩＡＡ０．２ｍｇＬ（－１）中生根成植株。可乐茄叶肉原生质体的植株再生,可应用于育种和茄属植物遗传工程研究。     

蝴蝶兰根段的组织培养

１　植物名称　蝴蝶兰（ＰｈａｌａｅｎｏｐｓｉｓＭｅｌｌｅｒＧｏｌｄ“ＮＦＳ”）。２　材料类别　根段。３　培养条件　（１）愈伤组织的诱导及分化培养基：Ｂ５＋ＮＡＡ１．５ｍｇ·Ｌ－１（单位下同）＋ＫＴ０．２＋ＣＭ１５０ｍｌ·Ｌ－１＋３％蔗糖;（２）原球茎增殖培养基：Ｂ５＋ＧＡ０．０５＋ＣＨ１２０＋３％蔗糖;（３）小苗生长培养基：１／２ＭＳ＋２０％香蕉泥＋２％蔗糖;（４）诱导生根培养基：１／２ＭＳ＋ＩＢＡ０．３＋２％蔗糖。上述培养基均加０．２％活性炭,０．５８％琼脂粉,ｐＨ为５．５;培养基在１２１℃高…     

大花蕙兰的快速繁殖技术研究

应用组织培养和快速繁殖技术对大花蕙兰开展了茎尖培养、试管苗微繁和成苗培育的研究,结果表明,有利于原球茎增殖的培养基为ＭＳ＋６ＢＡ１．０ｍｇ／ｌ＋ＮＡＡ０．５ｍｇ／ｌ＋２％～３％蔗糖;有利于原球茎分化的培养基为ＭＳ＋６ＢＡ０．４ｍｇ／ｌ＋ＮＡＡ０．２ｍｇ／ｌ＋２％～３％蔗糖。四年生以上试管苗可以开花,为其大规模工厂化育苗提供了科学的依据。     

石斛离体培养中ABA对诱导花芽形成的影响

由兰科植物铁皮石斛（Ｄｅｎｄｒｏｂｉｕｍ ｃａｎｄｉｄｕｍ Ｗａｌｌ．ｅｘ Ｌｉｎｄｌ．）种子诱导形成的愈伤组织,在光照下置于ＭＳ附加０．３ ｍ ｇ／ＬＮＡＡ 的培养基上繁殖,可以形成原球茎。将原球茎转入ＭＳ含２ ｍ ｇ／Ｌ ６－ＢＡ 和０．５ ｍ ｇ／ＬＮＡＡ 的培养基上,花芽形成频率为２７．０％ 。原球茎先在０．５ ｍ ｇ／ＬＡＢＡ的培养基上预培养１５ ｄ,再转入含２ ｍ ｇ／Ｌ６－ＢＡ 的ＭＳ培养基上培养,花芽形成频率明显提高,可达８４．４％ ,而且每株植株花的数目增加;但是在仅有ＡＢＡ 的ＭＳ培养基上培养的原球茎再生的植株未见花芽形成     

宫粉郁金的组织培养和快速繁殖

１植物名称宫粉郁金（Ｃｕｒｃｕｍａ　ｋｗａｎｇｓｉｅｎｓｉｓ）。２材料类别　块茎萌动芽。３培养条件　萌动芽生长培养基：（１）ＭＳ＋６－ＢＡ１ｍｇ·Ｌ－１（单位下同）＋ＮＡＡ０．２。不定芽增殖与愈伤组织诱导培养基：（２）ＭＳ＋６－ＢＡ１０＋ＫＴ５;（３）ＭＳ＋６．ＢＡ１０;（４）ＭＳ＋６－ＢＡ５＋ＫＴ２．５;（５）ＭＳ＋６－ＢＡ５;（６）ＭＳ＋６－ＢＡ２＋ＫＴ１。生根培养基：（７）ＭＳ＋ＮＡＡ０．５;（８）ＭＳ＋６－ＢＡ０．５＋ＮＡＡ０．５;（９）ＭＳ。以上培养基均加０．７％琼脂,３％蔗糖,ｐＨ５…     

墨兰的无菌播种和植株再生

墨兰的无菌播种结果发现,在不添加细胞分裂素的培养基上,种子可以发芽,但只有原球茎和根状茎产生,不可能进一步分化成苗,只有在含有不同激素成分ＭＳ或ＫｎｕｄｓｏｎＣ培养基上,才有可能诱导芽的分化,其中以附加６－ＢＡ０．５－１．０ｍｇ／Ｌ＿ＮＡＡ０．１ｍｇ／Ｌ诱导效果最佳,在附加６－ＢＡ２．０ｍｇ／Ｌ＋ＮＡＡ０．４ｍｇ／Ｌ的ＭＳ培养基中,能加速芽的增殖,根状茎转入含有相同激素成分的液体增殖培养基中振荡培     

大叶紫花苜蓿愈伤组织原生质体再生植株

大叶紫花苜蓿下胚轴诱导的愈伤组织在继代培养基上生长快速,易于分散。继代第１２ｄ的愈伤组织原生质体的得率为６．５×１０７／ｇ鲜重。原生质体培养基为ＳＨ基本培养基,含有１．０ｍｇ／Ｌ２,４－０、０．５ｍｇ／ＬＢＡ、２．０ｇ／ＬＣＨ、２％蔗糖、６％葡萄糖、５ｍｍｏｌ／ＬＭＥＳ,培养密度为１．０×１０５／ｍＬ。培养至第１２ｄ时的原生质体再生细胞植板率为３．７％。由原生质体形成的小愈伤组织在含２．０ｍｇ／Ｌ２,４－Ｄ的ＭＳ固体培养基上大量增殖。增殖的愈伤组织转移至２．０ｍｇ／Ｌ２－ｉｐ＋０．１ｍｇ／ＬＮＡＡ的Ｂ５培养基上,形成体细胞胚并发育成完整植株。     

紫羊蹄甲的组织培养和快速繁殖

１植物名称紫羊蹄甲（Ｂａｕｈｉｎｉａｐｕｒｐｕｒｅａ）。２材料类别带腋芽的茎段。３培养条件（１）启动培养基：ＭＳ＋６－ＢＡ２．０ｍｇ·Ｌ－１（单位下同）＋ＮＡＡ０．０１;（２）诱导分化培养基：ＭＳ＋６－ＢＡ１．０～２．０＋ＩＢＡ０～０．１;（３）生根培养基：１／２ＭＳ＋ＩＢＡ０．５～１．０。上述培养基均入蔗糖３０ｇ·Ｌ－１,琼脂０．５％,ｐＨ值５．８。培养温度２５℃,光照度２５００～３０００ｌｘ,每天光照１２ｈ。４生长与分化情况４．１启动培养材料取自南京中山植物园温室种植的三年生母树。将带腋芽的…     

真菌诱导子对象牙白原球茎生长的影响

在1/2MS基本培养基中,分别加入由野生春兰和华石斛根部分离到的内生真菌所制成的4种菌丝提取物,对象牙白种子萌发形成的原球茎进行诱导培养。结果表明:4种菌株诱导子中3种可不同程度地促进象牙白的生长和分化。对象牙白原球茎的诱导作用依次为HF26>CF6>CF8,与对照相比,HF26菌株显著提高了象牙白的鲜重(P<0.05),鲜重增长率分别为96.13%、86.86%和61.18%;干物质重分别增加了130.70%、96.50%和124.30%,均达到极显著水平(P<0.01)。此外,CF8菌株能促进根条数的增加;HF26菌株最有利于原球茎的增殖;CF6菌株在促进成苗和根原基的分化上效果显著。结论:在象牙白原球茎生长的不同阶段,能够促进其达到最佳生长状态的菌株是不同的。     

La^3＋对墨兰根状茎生长的调节作用

La~(3 )具有调节细胞膜透性,抑制糖和色素外渗、促进根对磷的吸收、促进根的发育等生理功能,但对其在植物生长和激素水平上调节作用的研究尚未见报道。本文以墨兰根状茎为材料,探讨了     

实验室条件下五唇兰菌根真菌专一性研究

利用从高原温带兰科植物菌根中获得的22个菌根真菌菌株, 对五唇兰(Doritis pulcherrima)进行了室内种子萌发、原球茎分化和组培苗回接试验, 从交叉回接的角度对附生兰科植物与菌根真菌的生理专一性进行了探讨。经过20周的共生培养, 只有编号为Cf1和Mm1的两个菌株使种子表现出种胚明显膨大的萌发迹象; 9个菌株能够促使原球茎较好地分化发育出根叶; 11个菌株处理苗的平均鲜重增长率高于对照组(156.25%), 其中Mm1的效果达到极显著水平(p = 0.01)。通过根切片显微观察, 在原球茎分化根和回接效果良好的处理苗的根皮层组织发现典型的菌丝团结构, 表明菌根体系已成功建立。温带地生兰菌根真菌对五唇兰种子萌发、原球茎发育和幼苗生长等3个重要生长阶段影响的试验显示, 五唇兰的种子和菌根真菌的共生萌发效果不佳, 而原球茎及幼株更容易与之建立良好的共生关系。同时, 也没有发现同一个真菌菌株能够对五唇兰的种子、原球茎和幼苗均产生促进作用。研究结果表明, 五唇兰的菌根真菌专一性因生理生长阶段的不同而存在差异。     

大花斑叶兰种子无菌萌发及快繁技术研究

对大花斑叶兰（Goodyera biflora）成熟种子无菌萌发、原球茎分化及根诱导进行了研究。结果表明：黑暗或弱光照培养有利于大花斑叶兰种子萌发,而较强的光照对形成健壮的原球茎有利;1/2MS＋NAA0.2mg/L培养基较为适合大花斑叶兰种子的萌发;MS＋KT1.0mg/L＋NAA0.5mg/L＋活性炭2.0g/L对原球茎增殖形成类原球茎有较好效果;1/2MS＋6-BA2.0mg/L＋NAA0.5mg/L＋2.0g/L活性炭对根诱导效果较为理想。     

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.     

EFFECT OF INITIAL SIZE ON GROWTH OF PLANT TISSUE CULTURES

Caplin , Samuel M. (LOS Angeles State Coll., Los Angeles, Calif.) Effect of initial size on growth of plant tissue cultures . Amer. Jour. Bot. 50(1): 91–94. Illus. 1963.—Using different slice thicknesses and cannula diameters, cylindrical expiants of different size were obtained from tubers of Jerusalem artichoke. Slice thickness ranged from 0.5 to 2.5 mm and expiant diameter from 1.30 to 3.22 mm. Initial fresh weights ranged from 2.3 to 29.8 mg, a 13-fold range. After 17 days growth at 26 C in 10 ml basal medium containing 15% coconut milk, relative increase was inversely related to initial weight and ranged from 36-fold for 2.3-mg expiants to 8-fold for 29.8-mg expiants, a range of 4.5 × in relative increase between the largest and smallest cultures. Secondary phloem expiants of the same diameter from carrot root slices of different thickness, all removed at 1 mm from the cambium, also showed decreased relative growth with increasing size of explant; this was true for each of the different diameters used. Further investigation showed clearly that the growth of the large inocula was not limited by the amount of culture medium used during the period of culture. Initial weight has, therefore, a decided effect on the rate of relative growth.     

Cultural requirements for in vitro seed germination, protocorm growth and seedling development of Geodorum densiflorum (Lam.) Schltr

Effects of different nutrient solutions, organic supplements and plant growth regulators on in vitro seed germination and protocorm development of Geodorum densiflorum (Lam.) Schltr. were studied. Seed germination was very high (up to 96%) in all the basal media, with Knudson's C and half-strength Murashige & Skoog being slightly more productive than Vacin & Went. Application of organic supplements and NAA had little effect on germination, but BAP proved inhibitory. After germination, protocorms exhibited a clear preference for peptone and NAA for much faster growth, while BAP resulted in stunted growth. Beside normal development, disorganisation of protocorms, followed by callusing occurred in presence of peptone and NAA. The calli were compact with limited growth and frequently regenerated protocorm like bodies. Development of seedlings was preceded by an intermediary rhizome phase. Growth of rhizomes was slow in the plant growth regulator free medium and about 15 months of culture was required for seedling formation. However, it was possible to hasten the process by 8-10 months with the employment of NAA, which also enhanced the number of seedlings per protocorm through axillary branching. Combined application of high BAP and low NAA was also useful for high rate of seedling formation.     

La3+对墨兰根状茎某些细胞器发育的影响

以１／２ＭＳ＋ＢＡ（１ｍｇＬ－１）＋ＮＡＡ（１ｍｇＬ－１）＋Ｌａ３＋（１０ｍｇＬ－１）培养墨兰根状茎３０ｄ后,取样观察根状茎细胞叶绿体、线粒体和细胞核的发育特点并与对照进行比较,发现：①处理组叶绿体发育较快,有较丰富的片层结构和嗜锇颗粒,而对照组的发育慢,含大型的淀粉粒,无明显的片层结构．②处理组线粒体体积较大,内含物较丰富,而对照组的较小,内含物较少。③处理组细胞核内的染色质较对照组丰富．上述结果表明,ｔａ３＋对墨兰根状茎细胞器的发育有一定的调节作用。     

春兰×大花蕙兰杂种试管苗开花现象

以野生春兰(Cymbidiumgoeringii)为母本,大花蕙兰(C.hybridium)为父本进行杂交,由杂交种子获得800多个春兰×大花蕙兰杂种原球茎无性系。其中一个杂种株系的原球茎继代培养2个月后,形成肉眼可见的花蕾。诱导花芽形成的最适激素组合为6-BA1.0mg/L+NAA0.1mg/L,总花芽形成率达31%。诱导花芽形成的最适材料为1￣2cm幼苗,花芽形成率达19%。     

花粉管通道法介导的铁皮石斛转基因技术

该研究以含有GFP和GUS基因的质粒和农杆菌为载体,采用花粉管通道法对铁皮石斛进行转基因技术研究。结果表明:(1)铁皮石斛种子萌发和原球茎生长对卡那霉素的最低致死浓度分别为90和150 mg·L~(-1)。进一步研究证实,在筛选转化种子和原球茎时,可分别向培养基中添加100和150 mg·L~(-1)的卡那霉素进行选择培养。(2)以携带GFP和GUS基因的质粒(pSuper1300和pBI121)和农杆菌为载体,用无菌去离子水重悬质粒pSuper1300和pBI121至浓度为100 ng·μL~(-1),用2%蔗糖+1/2MS+0.1%silwet-77+0.1%AS或5%蔗糖+0.1%silwet-77+0.1 mmol·L~(-1)AS重悬携带质粒pSuper1300和pBI121的农杆菌至菌液浓度为OD_(600)=0.7~0.8;在授粉后0.5~2.5 h内使用柱头滴加法导入携带外源基因的质粒或农杆菌溶液,收集成熟的转化种子,经选择培养及PCR检测发现,几乎所有处理的转化材料均能检测出外源GFP和GUS基因片段。另外,与农杆菌相比,以质粒为载体进行转化,可获得更高的结实率。该研究结果为铁皮石斛的基因工程育种提供了参考。