黄山药愈伤组织诱导与分化

采用黄山药野生植株作为外植体,试验了不同激素处理对黄山药愈伤组织的诱导、分化影响,结果表明：不同的外植体的诱导率差别较大,叶片的诱导率最高,最高达到85．7％,茎段的诱导率较低,平均诱导率仅10％左右。以叶片作为外植体诱导愈伤组织的最佳培养基配方为MS 2,4-D2．0mg／L 6-BA2．5mg／L;愈伤组织分化生芽的最佳配方为MS BA1．0mg／L NAA0．5mg／L 蔗糖2％ pH6．4;愈伤组织分化生根的最佳配方诱MS BA1．0mg／L NAA0．1mg／L 蔗糖3％ pH6．80。     

紫果猕猴桃幼胚愈伤组织诱导及植株再生

以紫果猕猴桃(Actinidia arguta var．purpurea)幼胚为外植体,诱导愈伤组织并进行植株再生。结果表明：不同的培养基和不同的培养条件对幼胚愈伤组织的诱导率及分化率不同;0．2mg／L ZT与0．5mg／L GA。配合使用有利于促进愈伤组织的诱导;7％蔗糖、600mg／L CH与400mg／L Gln都有利于促进愈伤组织的形成;在添加0．5mg／L 6-BA、0．05mg／L NAA与0．5mg／L GA3的MS培养基中植株的再生率达93．3％。     

芥蓝下胚轴离体培养及高频率植株的再生

三个芥蓝品种下胚轴离体植株再生的条件的研究结果表明：下胚轴切口处可直接诱导出芽,诱导“早花尖叶芥蓝”、“中花尖叶芥蓝”和“迟花尖叶芥蓝”直接出芽的最佳激素组合分别为2．0mgL-1BA,0．3mgL-1NAA＋2．0mgL-1BA,0．5mgL-1NAA＋2．0mgL-1BA,其相应的芽发生频率分别为84．6％,86．7％,93．3％。诱导芽发生的最适蔗糖浓度是1％。培养基中加入4．0mgL-1AgNO3和500mgL-1MES可显著提高芽再生频率。再生芽在MS附加0．1mgL-1NAA的培养基上诱导生根形成完整植株。离体再生苗与种子萌发实生苗田间生长外形差别不大,但长势稍慢。     

提高狗蔷薇离体培养植株再生频率

以狗蔷薇（Rosa canina Inermis）为材料,以MS为基本培养基,通过对不同植物生长调节剂的组合,大幅度提高了狗蔷薇离体培养植株再生频率。结果表明,2．0mg／L 6-BA＋0．3mg／L 2,4-D的组合较为适宜,其不定芽再生率为87％,增殖率为3．0;而CPPU和2,4-D的适宜组合为1．5mg／L＋0．3mg／L,其不定芽再生率高达93％,增殖率为5．0。同时,研究结果显示,以MS＋40g／L蔗糖＋6．0g／L琼脂粉＋3．5mg／L AgNO3＋1．5mg／L CPPU＋0．1mg／L 2,4-D＋0．05mg／L GA3作增殖培养基效果最好,不定芽诱导率为89％,增殖率为5．5;利于生根的培养基为1／4MS＋20g／L蔗糖＋3．5g／L琼脂＋0．3％活性碳＋0．1mg／L IBA＋0．1mg／L NAA,生根率为91％。     

提高西瓜离体培养植株再生效率的研究

本文以“京欣1号”母本和“伊选”西瓜4天苗龄子叶为外植体,研究离体培养植株高频率再生体系。结果表明：“伊选”子叶远轴端外植体的再生频率仅为10％,子叶近轴端外植体在5mg／LBA 0．1mg／L IAA的激素组合下植株再生频率为100％,平均每个外植体的丛生芽数在所有组合中最多,为10.3个;“京欣1号”母本子叶近轴端外植体在2mg／LBA 0．5mg／L IAA激素组合下植株再生频率为100％,平均每个外植体的丛生芽数在所有组合中最多,达6．9个。本试验条件下,子叶近轴端外植体接种4天即分化出不定芽,至再生苗的移栽仅需40天,在MS 0．1mg／L NAA的生根培养基上的生根率为97．3％,移栽成活率达98．5％。     

狗蔷薇茎尖再生体系的优化

采用正交设计及方差分析对影响狗蔷薇植株再生体系的因素进行了优化研究,结果表明:影响狗蔷薇植株再生的最主要因素是NAA和BAP,其次为AgNO3和蔗糖.适宜狗蔷薇茎尖再生的培养基为:MS+BAP1.5mg.L-1+NAA0.1mg.L-1+AgNO33.5mg.L-1+蔗糖40g.L-1+琼脂6.0g.L-1,植株再生频率高达96.3%,增殖系数为6.5.     

唐古特大黄组织培养技术的研究

试验选用唐古特大黄(Rheum tunguticum Maxim．ex Regel．)种了萌发的无菌苗及无菌苗子叶、下胚轴、胚根和幼根作为材料,研究唐古特大黄不同外植体的离体培养技术。结果表明,唐古特大黄的无菌苗和无菌苗子叶、下胚轴、胚根和幼根都可以作为离体培养的良好外植体。唐古特大黄的最适分化培养基足：B5 NAA0．1mg／L 6-BA3mg／L;最适乍根培养素是：1／2MS NAA1mg／L 3%蔗糖或1／2MS NAA0．5mg／L 3％蔗糖;愈伤组织诱导培养基是：MS 2,1-D 1mg／L NAA1mg/L 6BA1mg／L。     

影响早熟油桃胚轴培养及再生因素的研究

以早熟油桃华光、曙光为试验材料,对影响其胚轴再生的外源乍长调节剂浓度及组合、培养基、胚发育时期、低温处理等因素进行了研究。结果表明,胚发育时期在盛花后74d(74DAFB)更适合于做为胚轴再生的外植体。幼胚经过75d的低温处理,能够提高胚的萌发率,而且幼苗生长状态良好。胚轴能够直接再生植株。上、下胚轴无显著差异。华光油桃胚轴再生以G TDZ3．0mg／L KT1．0mg／L NAA3．0mg／L效果最好;曙光油桃胚轴再生以QL TDZ2．0mg／L NAA0．5mg／L效果最好。     

骆驼蓬的组织培养及植株再生

以骆驼蓬(Peganum harmala L)无菌苗下胚轴切段为材料,在不同的培养基上进行愈伤组织的诱导,发现在MS基本培养基附加2．0mg/L 2,4—D、0．5mg／L 6—BA和3％蔗糖时,可100％的诱导出愈伤组织。愈伤组织在附加2．0mg／L 6—BA、0．5mg／L NAA、500mg／L CH和3％蔗糖的MS培养基上诱导出丛生芽,进而发育成苗,苗的分化频率在30％左右。分化苗或其茎切断在附加0．2mg／L IBA、0．2mg/L NAA和3％蔗糖的l／2MS培养基上出现根的分化,分化频率在90％以上。再生植株经炼苗后移栽成活,成活率在80％以上。     

胡杨器官和体胚发生方式的植株再生

目的：为以胡杨为亲本的体细胞杂交育种奠定基础。方法：以胡杨苗叶片为外植体,通过器官和体胚两种不同发生方式建立了离体再生体系。结果：附加0．75mg/L BA、0．5mg/L NAA基本培养基及3w暗培养是愈伤组织诱导的最佳条件;附加0．25mg/L BA和0．1mg/L NAA的基本培养基上不定芽的诱导率最高;1／2大量元素的MS培养基附加0．1mg/l NAA、0．05mg／L和1．5％蔗糖对不定芽生根效果最好;诱导并筛选出的胚性愈伤组织在附加了0．5mg/L BA、0．5mg／L NAA的基本培养基上诱导获得大量胚状体,干化处理后大部分能经子叶胚期萌发成苗。结论：外植体的采集周期和培养条件影响胡杨离体叶片的形态发生途径。     

基因型和激素浓度对大豆植株再生的影响

本文采用8个大豆基因型,比较了NAA 激素浓度对大豆下胚轴植株再生的影响。结果表明：所有的基因型在合适的培养条件下, 均能形成愈伤组织,但不同基因型由愈伤组织诱导植株再生频率有很大差异。供试的8个品种中,在NAA浓度为0.1～0.5 mg／L的培养基上,汾豆33 号表现出较高的再生率,达50%;晋豆19号次之,为42%;而晋旱125则较低,仅19%。较合适的培养条件是：下胚轴在B5+NAA 0.3 mg／L+KT 1 mg／L+3%蔗糖,可高频率地诱导出愈伤组织并直接分化成苗。     

The Interaction of Genotype, Explant and Media on the Regeneration of Shoots from Complex Explants of Brassica napus L.

The relative importance of explant type, genotype and growthregulator regime in the determination of shoot regenerationfrequencies from complex explants of Brassica napus L. has beenevaluated. Cotyledon, hypocotyl and stem sections taken fromone spring (Westar) and three winter (Ariana, Cobra, Libravo)varieties of B. napus were cultured on three different growthregulator regimes, 0.5 mg dm^–3 NAA + 2.0mg dm^–3BAP, 0.5 mg dm^–3 NAA + 4.0mg dm^–3 BAP and 1.0mgdm^–3 NAA + 4.0mg dm^–3 BAP. The most significanteffects on shoot regeneration were due to explant type and variety.The regeneration from stem segments was not only two to threetimes higher than from hypocotyls or cotyledons, in all varieties,but the response was also more uniform across the varieties.The explant effect accounted for 44–95% of the regenerationresponse. In contrast, the contribution of growth regulatorregime was negligible. Although the growth regulator regimeas an independent effect was unimportant, regeneration fromboth Ariana and Libravo was significantly affected by the interactionof genotype with growth regulator regime. The importance ofboth the high shoot regeneration frequency from stem segmentsand the relative uniformity of response across the four testedgenotypes is discussed with respect to the potential benefitsof using this explant source in Agrobacterium-based transformationexperiments. Key words: Brassica napus, regeneration, genotype, tissue culture, complex explant     

改良菜心离体培养植株再生体系的研究（简报）

This investigation has developed an efficient and fast method for plant regeneration from petiole of cotyledon explants of Brassica campestris L. subsp. chinensis Makino var. parachinensis Tsen et Lee. A medium was designed for B. campestris subsp. chinensis var. parachinensis to obtain the high frequency of shoot regeneration, which contained BAP 2 mg/L, NAA 0.75-1.0 mgL and 7.5 mg/L AgNO3 solution to the half of NH4+ concentration's MS basic medium. 60 mL/L coconut milk were added to all of media. In this method, frequency of shoot regeneration of "youqing caixin" reached as high as 91.2% and the number of shoots per explant reached as high as 4.7 plants. The result showed that there was a positive correlation between frequency of shoot regeneration and number of shoots per explant. The little shoots could be observed five days after inoculation and were formed directly. The inducing rate of roots of the shoots reached as high as 100% and the rate of viability of transferred mature plant reached higher than 95%. The regeneration period from petiole with cotyledon to a seedling was shorten to about 49 days. Factors influencing in vitro explant regeneration were studied.     

An efficient in vitro regeneration system of fieldpea (Pisum sativum L.) via. shoot organogenesis

In-vitro regeneration in fieldpea was achieved from immature embryonic axes and cotyledonary node explants of six genotypes on modified MS media supplemented with different concentration of plant growth regulators, 6-Benzylamino purine (BAP) and Naphthalene acetic acid (NAA). The best regeneration response, leading to multiple shoot formation efficiency (22.34 shoots/explant) was observed in the medium supplemented with 1.0 mg/L BAP and 0.2 mg/L NAA and best frequency (67.55?±?4.74) was achieved on medium containing 2.0 mg/L BAP and 0.4 mg/L NAA. The shoots were subcultured on a medium supplemented with a combination of 1.0 mg/L GA₃, 2.0 mg/L BAP and 0.4 mg/L NAA, which resulted in elongation of 85 % of shoots. Rooting attempted from the elongated shoots, on half strength MS medium and supplemented with three different auxins IBA, IAA and NAA separately, exhibited similar results. Alternatively, micro-grafting of in vitro regenerated shoots onto pre-germinated root stocks raised in green house facility was attempted with high success rate (75 %). The grafted plants could be successfully hardened, fertigated with Hoagland solution and distilled water in a ratio of (1:10) for acclimatization and further development. All the genotypes tested, produced multiple shoots that could be established to mature fertile plant, hence, the medium combinations used were found to be genotype neutral.     

In vitro morphogenesis from cotyledon and epicotyl explants and flow cytometry distinction between landraces of Bambara groundnut [<Emphasis Type="Italic">Vigna subterranea</Emphasis> (L.) Verdc], an under-utilised grain legume

The morphogenetic competence of Bambara groundnut was assessed for different landraces, explant sources and media compositions. With cotyledon explants, the best callusing occurred on a medium containing 3 mg/l BAP + 0.5 mg/l NAA, while roots were produced with 3–5 mg/l BAP + 0.5 mg/l NAA. Shoots regenerated (∼6%) from cotyledons on media with BAP alone (3–5 mg/l) or combined with 0.01–0.1 mg/l NAA. Flowers were regenerated on 5 mg/l BAP + 0.5 mg/l NAA, without any intervening callus phase. With epicotyls, the highest callusing was on 3 mg/l BAP + 0.5 mg/l NAA, and shoots regenerated (15–20%) on 3 mg/l BAP alone or with NAA at concentrations that depended on the landrace studied. Regenerated shoots rooted on hormone-free medium, and plants transferred to the greenhouse were all morphologically normal and fertile. Flow cytometry showed that most regenerants were diploid and in addition permitted to distinguish between landraces according to their relative nuclear DNA content. This is the first report on de novo regeneration in vitro of Bambara groundnut, an important yet neglected legume crop.     

In vitro shoot regeneration from leaf explants of Echinops kebericho: an endangered endemic medicinal plant

Echinops kebericho is a critically endangered endemic medicinal plant of Ethiopia. It is threatened due to over harvesting of its roots for medicinal purposes and from poor seed viability. This study aimed to develop a protocol for in vitro shoot regeneration from leaf explants of E. kebericho. The seeds were sterilized using ethanol followed by Clorox or calcium hypochlorite. Shoots from the germinated seeds were cultured on Murashige and Skoog (MS) medium containing different concentrations of α-naphthalene acetic acid (NAA) and 6-benzyl amino purine (BAP). Young leaves were cultured on MS medium containing different concentrations of BAP and NAA for shoot regeneration. For shoot multiplication, shoots were excised and cultured on MS medium containing different concentrations of BAP or kinetin (KIN) and NAA. The highest mean number of initiated shoots (4.00 ± 0.57) with 100% shoot induction was obtained on medium containing 1.0 mg/L BAP and 0.2 mg/L NAA. The highest shoot regeneration (33%) and shoot number (2.13 ± 0.06) were obtained on MS medium containing 2.0 mg/L BAP and 0.5 mg/L NAA. Medium containing 1.0 mg/L KIN and 0.2 mg/L NAA produced the highest number of shoots (4.67 ± 0.33) per explant. This protocol can be used for genetic improvement and conservation of this endangered species.     

Somatic embryogenesis and plant regeneration from leaf callus of Ocimum basilicum L

A effective protocol for complete plant regeneration via somatic embryogenesis has been developed for Ocimum basilicum L. Callus was initiated from leaf explant of young plant on supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) 1.0 mg l(-1), 3% sucrose and 0.9% agar. The calli showed differentiation of globular structure embryos when transferred to MS medium containing 2,4-D 0.5 mg l(-1) and BAP 1.0 mg l(-1). The maximum globular structure embryos were further enlarged and produced somatic embryos in MS basal medium supplemented with BAP 1.0 mg l(-1)+NAA 1.0 mg l(-1) + KN 0.5 mg l(-1). Continued formation of globular embryo and germination of embryos occurred in this medium. Complete plantlets were transferred onto specially made plastic cup containing soilrite followed by their transfer to the garden soil. Survival rate of the plantlets under ex vitro condition was 80%.     

改良菜心离体培养植株再生体系的研究(简报)

菜心(Brassica campestris L.subsp.chinensis Makino var.parachinensis Tsen et Lee)为十字花科芸薹属芸薹种中国白菜亚种中的一个变种,又名“菜薹”。它是我国南方特产蔬菜之一,在蔬菜的周年供应上有重要地位。目前迫切需要培育抗病虫、抗逆和具有其他优良农艺性状的新品种,以提高菜心的     

采用俄罗斯橄榄叶片和茎段诱导产生不定芽的高效再生方法

俄罗斯橄榄(Elaeagnus angustifolia L.)是一种具有很重要药用价值和生态意义的植物。以俄罗斯橄榄一年生幼苗的叶片和茎段为实验材料,探讨了细胞分裂素类(6-BA和Zt)和生长素类(NAA和IBA)两类激素不同组合以及不同配比对植株再生的影响,最后建立了一个高效的俄罗斯橄榄再生方法。结果表明,MS 培养基+ 0.5 mg/L 6-BA +0.2 mg/L NAA更适合叶片的再生,平均每个外植体能产生多达4.3个不定芽;而在MS培养基 + 1.0 mg/L Zt +0.5 mg/L NAA的条件下,茎段外植体再生出来的不定芽最多可以达到平均3.6个;再生芽在含有0.5 mg/L NAA的1/2 MS培养基上生根率达到100%。体外再生苗移栽到装有灭菌混合土(土∶泥炭∶沙子=1∶1∶1)的花盆中锻炼驯化,最后有77%的再生植株存活下来。此结果不仅对俄罗斯橄榄种质资源保护有重要的促进作用,另外也为其将来的遗传转化奠定了基础。