桃幼胚下胚轴高频植株离体再生

以桃品种京艳和晚蜜不同发育阶段合子胚下胚轴为外植体进行离体培养,用6-BA、CPPU和TDZ等细胞分裂素诱导下胚轴直接再生不定芽试验。结果表明,花后70d下胚轴为诱导的最佳材料,供试细胞分裂素中,诱导活性为TDZ>CPPU>6-BA,苯脲类细胞分裂素TDZ5mg/L与NAA0.01mg/L配合对下胚轴再生具有极强的诱导作用,京艳诱导再生率高达95.2%,平均不定芽数达每外植体25.3个。京艳的诱导率显著高于晚蜜。IBA 0.5mg/L适于诱导再生幼茎的生根,生根率和移栽成活率分别达到92.6%和65.5%。组织学观察表明不定芽起源于皮层薄壁细胞,属直接再生方式。     

The effects of explant and cytokinin type on regeneration of Prunus microcarpa

We investigated in vitro regeneration ability of Prunus microcarpa subsp. tortusa using various explants (root, cotyledon and hypocotyl pieces) and cytokinins [benzyladenine (BA), meta-Topolin (mT) and thidiazuron (TDZ)]. Sectioned cotyledon, root and hypocotyl pieces of in vitro grown seedlings were cultured on Nas and Read Medium (NRM) containing BA (7.5, 10, 12.5, 15 or 17.5 μM), mT (2.5, 5.0, 7.5, 10 or 12.5 μM) or TDZ (2.5, 5.0, 7.5, 10 or 12.5 μM). As a measurement of morphogenetic reaction, the ratios of regenerating explants and the numbers of primary adventitious shoots per regenerating explant were analyzed. Cotyledon explants exhibited higher regeneration ratios than hypocotyl explants, and the root explants were inappropriate for regeneration. Both BA and mT were effective on shoot regeneration but higher regenerating explant ratios were obtained when BA was used. In comparison with BA and mT, the effect of TDZ on enhancing explant regeneration ability was insignificant. Mean number of adventitious shoot per regenerating explant was between 1 and 4, and regenerating explant ratios were between 0% and 77%. The practical appliacations of the results are discussed.     

蔓性蝴蝶草叶片的组织培养及植株再生

以蔓性蝴蝶草全展叶片为外植体,以MS为基本培养基,分别添加0.01、0.05、0.1 mg/L浓度的TDZ及CPPU,研究TDZ及CPPU对分化芽的影响,并进行了最佳生根培养基的筛选.结果表明:不同浓度的TDZ和CPPU对蔓性蝴蝶草的芽分化影响不同,MS+ TDZ0.1 mg/L和MS+CPPU 0.05 mg/L对外植体芽的分化的诱导效果最佳,达到100％;在1/3 MS培养基上的生根效果最佳;移栽成活率在90％以上.     

In vitro shoot regeneration from immature seeds of Epimedium alpinum induced by thidiazuron and CPPU

In vitro propagation of Epimedium alpinum L. was carried out using immature seed explants. The effects of various concentrations of thidiazuron (TDZ) and 1-(2-chloro-4-pyridyl)-3-phenylurea (CPPU), on the induction of organogenic callus, were evaluated. Organogenesis occurred most efficiently when explants were transiently exposed (48 h) to 20 μM CPPU or 80 μM TDZ followed by culture on hormone-free woody-plant medium (WPM). Organogenic callus consisting of white, compact clumps of tissue proliferated slowly on hormone-free WPM. To promote adventitious shoot induction, the effects of different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine (BA) were investigated. The highest per cent shoot regeneration, 66.7% of explants, and the maximum mean number of shoots, 2.6 per explant, were obtained on WPM containing 1.1 μM 2,4-D and 22 μM BA. Shoots were rooted on hormone-free WPM and well-developed plantlets were successfully transferred to soil.     

Adventitious bud formation and plantlet regeneration from leaves of fig (Ficus carica L.)

Summary

Leaf fragments of fig (Ficus carica L. ‘Masui Dauphine’) regenerated from in vitro shoot culture were excised and inoculated on MS medium supplemented with different combinations of 2, 4-D, TDZ, and 0.5 mM phloroglucinol. Addition of 2, 4-D induced root formation directly on the explant, and the presence of phloroglucinol significantly increased root formation. When a combination of 2, 4-D and TDZ was added to MS medium containing phloroglucinol, the explants started to produce adventitious buds at the edges. The addition of phloroglucinol was effective in inducing adventitious bud formation. Excised shoots were rooted successfully in MS medium that was either hormone free or supplemented with 1.0 mg l^–1 indolebutyric acid. Regenerated plantlets were successfully established in soil after a short period of acclimatization. This is the first protocol of organogenesis and plant regeneration from vegetative organs of Ficus carica L.     

茎段外植体白化处理促进“皇家嘎拉”苹果不定芽再生

研究比较了来自正常和白化处理的皇家嘎拉（Royal Gala)苹果（Malus domestic Borkh)新梢节段外植体不定芽的离体再生，。结果表明，白化处理的新梢顶部第一节段外植体所产生的不定芽数比第二、第三、第四节段分别高2、8、73倍；白化新梢的外植体不定芽再生数目比同一部位正常新梢高7倍。培养基添加较低浓度的TDZ有利于不定芽再生，对白化外植体来说，1μmol/L比10μmol/L高2．5倍，非白化外植体则高7倍,平均每个外植体的不定芽再生数高达68．8个。白化第一节段外植体的不定芽再生能力比常用的叶片外植体高4倍，表明该类外植体可用于苹果的转基因和染色体组工程育种。     

Effects of media,carbon sources and cytokinins on shoot organogenesis in the Christmas tree Scots pine (Pinus sylvestris L.)

Summary

Significant effects of seven basal media and three carbon sources (sucrose, glucose and fructose) on the induction of adventitious buds from embryos of Pinus sylvestris L. were observed. Moreover, hyperhydricity of expiants and shoot regenerants was observed on basal media containing fructose, especially with half-strength Murashige and Skoog (MS), MS, and woody plant medium (WPM). Expiants grown on a Gresshoff and Doy (GD) medium with sucrose produced the highest frequency of regeneration (81%) and with no hyperhydricity observed of developing adventitious shoots. Among three cytokinins tested including BA, BPA, and TDZ (at four concentrations each), 5 μM BA resulted in the highest regeneration frequency and mean number of adventitious shoots per embryo. Shoot régénérants were elongated after transfer to a GD medium containing 2 g-l_–1 activated charcoal and no growth regulators. After one month, rooting was induced on 10% of expiants.     

An improved protocol for adventitious shoot regeneration and plant formation in Psoralea corylifolia L.

An effective protocol was developed for in vitro regeneration of Psoralea corylifolia through enriched cotton moistened-liquid (CML) and solid culture systems. Prolific adventitious shoot buds were achieved from hypocotyl explants of 2-week-old cultures on enriched CML Phillips and Collins (L2) medium supplemented with different concentrations and combinations of thidiazuron (TDZ), benzyladenine (BA), kinetin (KIN), naphthalene acetic acid (NAA), indole-3-acetic acid (IAA) and bavistin (BVN). Combination of 2 μM TDZ, 0.5 μM BA, 100 mg l⁻¹ BVN and 2 μM NAA produced a greater number of adventitious shoots per explant (93.5) when transferred to half-strength enriched solid L2 medium. Regenerated shoots (40–50 mm in length) were exposed simultaneously for rooting as well as hardening in moistened (1/8-L2 basal salt solution with 5 μM IBA and 100 mg l⁻¹ BVN) soil mixture and vermiculite (3:1, v/v). The plants were subsequently established in the field. The survival percentage differed with seasonal variations.     

美人指葡萄不定芽离体诱导再生植株的研究

6-BA、TDZ与IBA不同浓度组合对美人指葡萄叶片、叶柄和茎段不定芽离体器官发生途径诱导再生植株有不同的影响。以茎段为外植体在MS+6-BA1.0mg/L+IBA0.01mg/L培养基上分化效果最好,再生率75.00%;叶片、叶柄分别在MS+6-BA3.0mg/L+IBA0.10mg/L和MS+6-BA2.0mg/L+IBA0.05mg/L培养基上分化效果较好。TDZ和IBA研究的几种组合都能诱导不定芽再生,但再生率不高。     

Regeneration of plants from Fraxinus pennsylvanica hypocotyls and cotyledons

An adventitious shoot regeneration and rooting protocol was developed for green ash (Fraxinus pennsylvanica) seedling explants. The best regeneration medium for freshly isolated hypocotyls and cotyledons was Murashige and Skoog (MS) supplemented with 13.3 μM 6-benzylaminopurine (BA) plus 4.5 μM thidiazuron (TDZ), and 22.2 μM BA plus 4.5 μM TDZ, respectively. Seventy-six percent of hypocotyl segments and 24% of cotyledon segments produced adventitious shoots, with a mean number of adventitious shoots per explant of 2.7 ± 0.5 and 2.3 ± 1.3, respectively. The effect of in vitro-germinated seedling age on adventitious shoot regeneration from hypocotyl and cotyledon explants was also studied. Results showed that hypocotyl and cotyledon explants from freshly isolated embryos exhibited a higher organogenesis potential than 4–15-day-old explants. Adventitious shoots from hypocotyls and cotyledons were established as proliferating shoot cultures following transfer to MS basal medium with Gamborg B5 vitamins supplemented with 10 μM BA plus 10 μM TDZ. A high rooting percentage (73–90%) was achieved when in vitro shoots were rooted on woody plant medium (WPM) containing 4.9 μM indole-3-butyric acid (IBA) and IAA (0, 2.9, 5.7, or 8.6 μM) with a combination of 10-day dark culture period followed by a 16-h photoperiod. The highest rooting (90%) of adventitious shoots or the number of roots per shoot (3.0 ± 1.0) was obtained on WPM with 4.9 μM IBA plus 5.7 μM IAA. Rooted plants were successfully acclimatized to the greenhouse and 100% survived after overwintering in cold storage. This regeneration system using hypocotyls and cotyledons provides a foundation for Agrobacterium-mediated genetic transformation of F. pennsylvanica for resistance to the emerald ash borer.     

Rapid in vitro multiplication and conservation of Garcinia indica: A tropical medicinal tree species

A simple and efficient method has been developed for rapid regeneration of plantlets via adventitious bud differentiation on mature seeds of Garcinia indica (Thouars) Choisy, a medicinally important facultative apomictic tropical tree species. High frequency direct shoot proliferation was induced in seed segments cultured on Murashige and Skoog's medium supplemented with cytokinins (BAP, kinetin and TDZ) alone and in combination with auxin (NAA). Amongst the various combinations used, BAP proved to be the most effective. Multiple shoots formed within 4–5 weeks of culture. The shoot forming capacity of the seeds was influenced by the BAP concentration tested (5–50 μM) and optimal response was observed at different concentrations (12.5–50 μM) in different genotypes investigated. Significant differences were recorded in terms of percent response (27.78–100%) as well as average number of shoots per explant (3.49–57.67) among the four genotypes investigated. Elongation of the induced shoots was achieved on MS basal medium containing 0.2% activated charcoal. The induction medium had a profound effect on rate of bud elongation with shoots induced on lower concentrations of BAP showing as much as four-fold elongation within 4 weeks. Proliferating shoot cultures were established by repeatedly subculturing the shoot nodes on MS medium supplemented with 5 μM BAP. Maximum rooting (91.66%) occurred in shoots cultured on half-strength MS medium supplemented with 10 μM IBA. The plantlets were successfully transferred to soil after hardening; the time period from initiation of shoot buds to transplantation being 15–17 weeks. For in vitro conservation, the shoot cultures were maintained on medium supplemented with 0.5 μM BAP and the subculture duration could be enhanced up to maximum of 11 months.     

欧李离体叶片再生体系的建立

【目的】以欧李试管苗叶片为外植体进行不定芽再生研究,以期为核果类果树的品种改良、基因遗传转化和功能验证等生物技术育种奠定一定的基础,【方法】使用不同的基本培养基和激素组合促进其不定芽再生并建立再生体系。【结果】结果表明,欧李幼芽增殖对基本培养基类型较敏感,改良MS培养基优于MS、F14和WPM。在MS(改良)+NAA 0.1 mg.L-1+BA 0.2 mg.L-1培养基上,增殖系数为5.6,增殖效果较好。离体叶片不定芽再生的最适植物生长调节剂组合为MS(改良)+TDZ 4.0 mg.L-1+IBA 0.2 mg.L-1,再生率达到63.1%,平均每外植体再生芽数为4.9。暗培养10 d可以提高不定芽再生率。再生不定芽1/2 MS(改良)+IBA3.0 mg.L-1培养基上生根率最高,生根率达93.3%。【结论】在改良MS培养基上TDZ与IBA组合诱导不定芽再生的效果最好。     

中国樱桃‘对樱桃’不定根离体再生植株的研究

 采用中国樱桃‘对樱桃’( Prunus pseudocerasus) 无菌生根苗的不定根作为外植体, 成功诱导了胚性愈伤组织, 实现了通过分化不定芽和诱导初生体细胞胚两种方式再生植株。根切段在MS + NAA1.0 mg/L + BA 0.05 mg/L + IBA 0.05 mg/L 培养基诱导获得胚性愈伤组织, 诱导频率达54.2 % , 该愈伤组织在MS + NAA 1.0 mg/L + BA 0.5 mg/L 培养基上不定芽分化率为100 % , 不定芽在MS + BA 0.5 mg/L + IBA 0.1mg/L 培养基上生长发育良好, 转入MS + NAA 0.02 mg/L + IBA 0.02 mg/L 生根培养基生根率达100 %; 整体不定根系统在MS 附加2 ,4-D 1.0～3.0 mg/L 和BA 0.5 mg/L 的培养基上同时诱导初生体细胞胚发生和单极不定芽发生, 每外植体上平均体细胞胚数5～25 个, 不定芽3～22 个。体细胞胚转到MS + BA 0.5 mg/L +IBA 0.1 mg/L 培养基上发育为完整植株, 植株再生率100 %。     

植物生长调节剂对珠芽黄魔芋叶柄离体培养效果的影响

以珠芽黄魔芋无根组培苗的叶柄为外植体材料,通过正交实验法研究了植物生长调节剂对其愈伤组织诱导、不定芽诱导、继代增殖的影响并进行生根试验,以期建立珠芽黄魔芋叶柄离体快繁体系。结果表明:2,4-D、6-BA及NAA对愈伤组织诱导率的影响差异达极显著水平(P<0.01),影响效应主次顺序为2,4-D>NAA>6-BA,优组合为2,4-D 0.5 mg·L-1+6-BA 1.0 mg·L-1+NAA 0.4 mg·L-1;IAA、6-BA、GA3及TDZ对平均不定芽数的影响达极显著水平(P<0.01),影响效果主次顺序为TDZ>6-BA>IAA>GA3,优组合为IAA 0.6 mg·L-1+6-BA 1.0 mg·L-1+TDZ 0.02 mg·L-1+GA30 mg·L-1,平均不定芽数为4.88个,生长良好;6-BA及GA3对不定芽增殖系数的影响达极显著水平(P<0.01),而IAA的影响不显著(P>0.05),影响效应主次顺序为GA3>6-BA>IAA,优组合6-BA 2.0 mg·L-1+IAA 0.6 mg·L-1+GA30 mg·L-1,增殖系数高达4.54;单独附加NAA 0.5 mg·L-1的1/2MS利于不定根的分化,生根率高达99.33%,平均根数为8.20条,平均根长为3.37 cm,根系效果指数为0.046。     

Morphogenetic responses of Salpiglossis sinuata L. leaf explants and callus

Leaf explants of Salpiglossis sinuata L. were cultured on MS medium containing the auxins 2,4-D, IAA, NAA, or the cytokinins 6-BAP, 2ip, or K singly or NAA + 6-BAP combinations (0.01 to 10.0 mg/l) to determine their morphogenetic responses. IAA and NAA induced callus or roots at all levels except IAA at 0.01 mg/l. Callus varying in friability was obtained on MS + 2,4-D at various concentrations. Friable, uniform dividing callus was obtained on UM medium. Optimum adventitious shoot formation occurred on leaf sections and from callus cultures placed on MS + 2ip. Rooting-difficulties were encountered, but 75% rooting efficiency was obtained on shoots cultured in MS + 0.001 mg/l 2,4-D. Rooted plants were readily transferred to the greenhouse, and flowered.     

扭果花组织培养及快速繁殖的研究

以扭果花试管苗叶片为外植体,MS为基本培养基,添加不同浓度的6-苄基腺嘌呤（6-BA）与萘乙酸（NAA）,以此来探索这两种植物生长调节剂对扭果花叶片诱导不定芽的影响。结果表明,将外植体接种在MS＋BA0．5mg／L＋NAA0．5mg／L＋蔗糖30g／L＋琼脂9g／L上不定芽诱导率最高;不定芽继代增殖的最适培养基为MS＋BA2．0mg／L＋NAA0．2mg／L＋蔗糖30g／L＋琼脂9g／L;生根效果最佳培养基为MS＋NAA0．1mg／L＋蔗糖50g／L＋琼脂9g／L,生根率达100％。     

铁线莲‘Multi-Blue’不定芽及体细胞胚发生的初步研究

 用‘Multi-Blue’铁线莲的幼叶、茎尖、嫩茎作为外植体进行培养，发现外植体来源以茎尖为最优，基本培养基以低盐的1/2MS为最佳。茎尖在附加0.5 mg·L-1TDZ+0.01 mg? L-1NAA+2%蔗糖的1/2MS固体培养基上培养1个月后芽增殖率达200.8%，且幼苗长势健壮。茎尖在附加0.8 mg· L-1TDZ+0.5 mg· L-1NAA+2%蔗糖的1/2MS固体培养基上培养时可形成淡黄色块状至颗粒状愈伤，经多次继代培养后可逐渐形成胚性愈伤组织，胚状体诱导率可达60%，进而获得再生植株。     

Direct somatic embryogenesis and adventitious shoot formation from immature axillary buds of Melastoma affine

Summary

Direct somatic embryogenesis and adventitious shoot formation were induced from immature axillary bud explants of Melastoma affine cultured on induction medium containing both naphthaleneacetic acid (NAA) and cytokinins. Among the cytokinins tested, thidiazuron (TDZ) played a greater role in the induction of somatic embryogenesis than 6-benzylaminopurine (BA) and kinetin (KT). Histological studies of paraffin sections showed that the same tissue from immature axillary buds produced different types of explants that developed floral primordia and vegetative bud primordia during the earlier and later flowering stages, respectively. These could result in different developmental pathways. Efficient mass propagation and plant regeneration systems were established for Melastoma affine.     

高品质甜叶菊品系“1096”的组培快繁

采用改良DNS等方法筛选出的生长快、糖苷含量高的甜叶菊品系＂1096＂为材料,研究了不同外植体、不同激素种类和配比对愈伤组织诱导、不定芽分化和生根的影响。结果表明：以茎尖为外植体愈伤组织和不定芽诱导率均最高;其次是带腋芽茎段;再次为叶片。无腋芽茎段和无菌根均能诱导出愈伤组织,但不能诱导成芽。茎尖不定芽诱导的最佳培养基为MS＋6-BA 1.0mg/L;叶片不定芽诱导的最佳培养基为MS＋6-BA 1.0mg/L＋IAA 1.0mg/L;带腋芽茎段不定芽诱导的最佳培养基为MS＋6-BA 1.5mg/L。不定芽继代增殖培养采用MS＋6-BA 0.5mg/L＋NAA0.05mg/L;在1/4MS＋IBA 0.1mg/L＋NAA 0.1mg/L＋1g/L活性炭的培养基上诱导生根,生根率可达100%,生根苗移栽后成活率达95%。     

不同细胞分裂素对2001富士苹果离体叶片再生的影响

以综合经济性状优良的2001富士苹果为试材,研究了不同浓度的细胞分裂素对其离体叶片再生效率的影响。结果表明:2001富士离体叶片分化不定芽适宜的BA、CPPU和TDZ的最佳再生浓度范围分别为8.0mg/L、6.0~8.0mg/L和1.0~2.0mg/L,再生频率分别达到67.9%、78.6%~89.3%和100.0%,ZT和KT不适宜诱导2001富士叶片分化不定芽。