我国藏红花离体快繁技术研究进展

藏红花离体快繁技术是解决我国藏红花资源短缺、繁殖系数低及球茎退化问题的有效途径。研究综述近几年我国藏红花离体快繁技术研究进展,总结并探讨藏红花离体快繁技术研究结果,为建立高效稳定的藏红花离体快繁体系提供参考。     

草莓品种石莓7号的组织培养与离体快繁

以草莓品种石莓7号茎尖为外植体,添加不同生长素的MS培养基,对其进行组织培养研究。结果表明:草莓茎尖在MS+BA 1.5 mg/L+NAA 0.1 mg/L培养基上能很好地诱导不定芽形成,诱导率95%;在继代增殖培养基MS+BA 0.5 mg/L+IAA 0.05 mg/L中增殖系数较高,为3.5;生根培养基为1/2MS+IBA 0.3 mg/L+IAA 0.3 mg/L,生根率可达95%;将生根瓶苗炼苗一周,移植至装有细土:蛭石(2∶1)的营养钵内,移栽试管苗的成活率达90%以上,并且生长健壮。     

萨米特甜樱桃离体快繁技术研究

以萨米特甜樱桃1年生枝休眠芽为外植体,进行离体快繁技术研究.结果表明,最佳诱导培养基为MS 6-BA 1.0mg/L GA0.5mg/L;最佳增殖培养基为改良MS 6-BA 1.0mg/L IBA 0.25mg/L;最佳生根培养基为1/2MS IBA 0.5mg/L.     

Rapid vegetative propagation of Hyacinthus orientalis L. in vitro

A simple in vitro system was developed to increase the vegetative propagation rate of hyacinth, Hyacinthus orientalis L. Excised bulb scale segments were induced to regenerate bulblets on the basal sides of the inverted explants. Within 12 weeks, 240–300 bulblets were obtained from a single 17–18 cim bulb when cut into basal scale segments, 3–4 cm long and 0.5 cm wide.     

Influence of medium composition and vessel ventilation on in vitro propagation of Phillyrea latifolia L.

Micropropagation of Phillyrea latifolia L. a wild species present in Mediterranean coastal areas having drought and salt tolerance was performed using explants from adult plants. Shoots were induced from nodal explants on the Rugini’s initial medium (IM). Then these were proliferated on either Rugini olive medium (OM) or Linsmaier and Skoog (LS) medium, each supplemented with 2.22 μM 6-benzylaminopurine (BA) or 4.56 μM zeatin (Z). Rooting (66.1±11%) was induced on shoots grown in perlite soaked with half-strength Rugini olive proliferation medium (OMr) containing 2.69 μM α-naphthaleneacetic acid (NAA) and 160 mg l⁻¹ putrescine. Both shoot multiplication and rooting were performed using Magenta^® GA-7 (Sigma) vessels either non-permeable or permeable to gas exchanges. Contamination (about 40%) was observed during the first five passages notwithstanding the addition of cefotaxime to the culture medium, but a high proliferation rate (90%) of explants provided enough healthy plant material. The highest shoot proliferation was observed on LS medium and zeatin whereas the presence of the ventilated filters reduced fresh weight of explants growing on LS media and did not affect shoot growth on OM media. During rooting, the use of ventilated vessels in comparison with the closed ones enhanced development of roots, and doubled the dry weight of plantlets. The vessel ventilation combined with the artificial substrate (perlite) was beneficial for in vitro acclimatization of rooted Phillyrea plantlets.     

西葫芦组培苗驯化和移栽影响因素分析

以西葫芦离体雌核发育而成的胚囊植株组培苗为试材,分析影响西葫芦组培苗驯化、移栽成活率的若干因素。结果表明,开始驯化时组培苗的苗龄、驯化温度、移栽基质组成、驯化方式等对西葫芦组培苗的驯化、移栽有较大影响;7～8叶1心的组培苗,在昼温22～28℃、夜温12～18℃的变温条件下,先闭瓶适应自然光照再开瓶逐步适应自然湿度,采用蛭石∶草炭=1∶1的移栽基质配方,成活率在95.0%以上。     

Effects of nutrients on the growth and survival of in vitro Artocarpus heterophyllus Lam. plantlets after transfer to ex vitro conditions in the glasshouse

Summary

The effects of nutrients in the planting substrate on the growth and survival of in vitro jackfruit (Artocarpus heterophyllus Lam.) plantlets under ex vitro conditions was investigated. A low level or absence of nutrients in the substrate for at least 20 days resulted in significantly more growth and survival of plantlets. Nutrient feeding after 20 days improved the further growth and survival. Nitrogen, particularly in the ammonium form, was shown to be the most inhibitory nutrient during the early stages of establishment.     

Screening of vincristine yield in ex vitro and in vitro somatic embryos derived plantlets of Catharanthus roseus L. (G) Don.

Catharanthus roseus L. (G.) Don. (Apocynaceae) is an important dicotyledonous medicinal plant as it is the sole source of vincristine and vinblastine that are used against a variety of cancers. Quantitative estimation of vincristine was carried out using high performance liquid chromatography (HPLC) in various in vitro grown tissues; calluses (embryogenic and non-embryogenic callus), embryogenic stages (proliferated, matured and germinated embryos), somatic embryo derived plantlets (leaf, root and whole plant) and leaves of ex vitro developed plantlets. The yield in those in vitro and ex vitro-developed tissues was monitored for 30 weeks. Except at an early lag period, vincristine production was detected up to 20–25 weeks old plant samples. Vincristine content was very high in leaf callus and germinated embryos. Leaves of in vitro raised plants showed higher amount of vincristine when compared to ex vitro-developed leaves of similar age. Vincristine production was tissue specific and age dependent that was discussed in detail in this present communication.     

Rapid in vitro propagation of Dendrobium hybrids through direct shoot formation from foliar explants,and protocorm-like bodies

In vitro propagation protocol for Dendrobium hybrids Sonia 17 and 28, two highly priced commercial cut flower cultivars through direct organogenesis from in vitro derived foliar explants was established. Rapid clonal propagation was achieved by subsequent induction of protocorm-like bodies (PLBs) and its conversion to shoots. No significant differences were observed in the induction of direct shoots, shoot multiplication, PLBs formation and subsequent shoot development and rooting of shoots between the two cultivars. Leaf explants from flower stalk node derived shoots cultured on half-strength Murashige and Skoog (MS) medium supplemented with 44.4 μM N⁶-benzyladenine (BA) developed more than seven shoots per explant. The isolated shoots transferred onto the same medium induced more than eight PLBs from the base within 60 days, which upon transferral to fresh medium having the same level of BA facilitated rapid proliferation. More than 200 PLBs were yielded from fifth subculture. Half-strength MS medium containing 6.97 μM kinetin (Kn) facilitated conversion of more than 90% PLBs to shoots. PLBs exhibited proliferation without decline up to the 15th subculture. Half-strength MS medium with 2 g l⁻¹ activated charcoal was the best for in vitro rooting. Plantlets of the hybrids exhibited more than 80% ex vitro establishment.     

杨桃SCoT标记PCR反应体系建立与验证

用单因素设计法对影响杨桃SCoT-PCR反应体系的主要因素Mg2+、dNTPs、引物、Taq DNA聚合酶及DNA模板浓度进行优化。结果表明,20μL反应体系中,含Mg2+2.5 mmol/L,dNTPs 0.3mmol/L,模板DNA 30mg/L,引物1.00μmol/L和Taq DNA聚合酶0.4U为最佳反应体系。用不同引物及杨桃DNA对该体系进行验证,扩增条带清晰,结果稳定可靠,证明该反应体系适用于杨桃SCoT-PCR扩增。     

Effect of benzyladenine, 4-PU-30 and thidiazuron on millisecond delayed and prompt chlorophyll fluorescence of Dianthus caryophyllus L. axillary buds cultured in vitro

The effects of benzyladenine (BA) and two phenylurea cytokinins (N-phenyl-N′-(2-chloro-4-pyridyl)urea (4-PU-30) and thidiazuron (TDZ)) on in vitro cultures of carnation were studied using induction kinetics and thermograms of prompt and delayed chlorophyll fluorescence. In plantlets obtained from explants cultivated on a medium with 0.4 μM BA or 0.04–0.004 μM phenylurea cytokinins, the photosynthetic apparatus showed high PS II photochemical activity comparable to control without cytokinins and 5.8–97.5% higher estimated photoinduced transmembrane proton gradient (low concentrations of 4-PU-30 are especially effective). At a concentration of 0.4 μM, when compared to BA treatment, phenylurea cytokinins induced 24–35% suppression of photochemical activity, 44–46% suppression of estimated transmembrane proton gradient and 12–17% suppression of estimated electron transport rate. It is likely that this concentration of phenylurea cytokinins exceeded the optimal dose. The analysis of the same luminescent characteristics in the presence of different 4-PU-30 and TDZ concentrations showed that these compounds induced dose-dependent changes in the functional activity as well as alteration in the level of energization of the thylakoid membrane. When fluorescence thermograms were evaluated, phenylureas appeared to change the high-temperature stability of the photosynthetic apparatus.     

Effects of chlorocholine chloride and prohexadione-Ca on rhizome growth and lateral bud production in Iris germanica L.

Iris germanica L. is a popular perennial flower worldwide, but its use is limited in China due to an inadequate availability of propagules. To accelerate rhizome growth and lateral bud production using plant growth retardants (PGRs), chlorocholine chloride (CCC; at 1,500 or 3,000 mg l^?1) or prohexadione-Ca (at 700 or 1,500 mg l^?1) were applied to uniform plants of I. germanica. Time-course measurements of changes in morphogenesis, carbohydrate metabolism, total soluble protein (TSP) concentrations, and endogenous phytohormone concentrations in rhizomes were conducted to test the efficacy of CCC or prohexadione-Ca for increasing rhizome growth and lateral bud production. The results showed that both PGRs increased the fresh weights of rhizomes at 2, 4, and 6 weeks after treatment (WAT). Overall, 700 mg l^?1 prohexadione-Ca was most effective at promoting the formation of lateral buds which increased by 183.5% at 12 WAT relative to untreated control plants. Concentrations of sucrose and starch in PGR-treated rhizomes increased at 2, 4, and 6 WAT, while a decline was observed by 12 WAT. TSP concentrations increased during rhizome enlargement, then decreased during lateral bud germination after prohexadione-Ca treatment. In general, concentrations of endogenous phytohormones, such as gibberellins, indole-3-acetic acid, jasmonic acid, and zeatin riboside, decreased significantly in rhizomes at 4 WAT, then increased at 12 WAT. Our study indicated that prohexadione-Ca promoted rhizome growth and the accumulation of sucrose and starch before summer dormancy, then significantly accelerated the production of lateral buds.     

杜梨组培生根过程中多胺、内源激素及相关氧化酶活性的变化

【目的】探究杜梨组培苗生根诱导过程中内源激素、多胺类物质及相关氧化酶活性变化的生理响应机制。【方法】以长势相近的杜梨继代苗为试材,筛选优化了生根培养基配方,分别在生根诱导0、3、7、10、15 d后,测定分析了基部茎段多胺类物质、内源激素含量及关联酶活性。【结果】(1)激素配比2.0 mg·L~(-1)IAA+0.5 mg·L~(-1)IBA的生根效果良好,IAA和IBA两种激素共同诱导杜梨生根效果要优于单一激素处理;(2)在生根诱导0～15 d内,精胺(Spm)和亚精胺(Spd)含量呈现下降→升高→下降→升高的变化趋势,腐胺(Put)含量则呈现出下降→上升的变化趋势;内源激素IAA、ABA含量均呈现先下降后升高的趋势,最低值出现在第7天,GA含量也呈现先下降后上升的变化趋势,最低值出现在第3天,ZR含量呈现出下降→升高→下降的变化趋势;两种关联酶IAAO和PPO酶活性均呈现增高→降低→增高的趋势(;3)Spd与IAA、ABA、IAA/ABA IAA/ZR及IAAO酶活性均存在极显著正相关关系(p 0.01),IAA与Spd、GA、ABA及IAA/ABA、IAA/ZR、PPO酶活性在α=0.01水平达到极显著正相关。【结论】杜梨较适宜的生根培养基为:1/2MS+2.0 mg·L~(-1)IAA+0.5 mg·L~(-1)IBA+7.5 g·L~(-1)琼脂+25.0 g·L~(-1)蔗糖;IAA/ABA、IAA/ZR比值增大更有利于杜梨不定根的发生;多胺类物质中Spd含量与杜梨不定根发生紧密相关;生根诱导后期,GA含量增加对杜梨不定根发生并无明显的抑制作用。     

Clonal propagation of mulberry (Morus indica L. cultivar M-5) through in vitro culture of nodal explants

A high frequency of sprouting (80.0%) and shoot differentiation was observed in the primary cultures of nodal explants of Morus indica L. cultivar M-5 on MS medium supplemented with 2,4-D (0.3 mg/l). In vitro proliferated shoots were multiplied rapidly by culture of shoot tips on MS medium with BAP (0.5 and 1.0 mg/l) which produced the greatest multiple shoot formation. Multiplication was also achieved by culture of shoot tips on MS medium with BAP (4.0 mg/l) and GA₃ (0.05 mg/l) which facilitated the elongation of shoots followed by sprouting of axillary buds of in vitro grown shoots. A high frequency of rooting (86.7%) with development of healthy roots was observed from shoots cultured on medium with 2,4-D (1.0 mg/l). Plants with well developed roots were transferred to soil with a survival frequency of 80%.     

苦瓜子叶离体再生体系的建立

以“圣香”苦瓜、“玉华”苦瓜的子叶为外植体进行离体培养植株再生研究.结果表明:2个苦瓜品种的子叶在不同的培养基上都较易形成愈伤组织,愈伤组织诱导率都达到80％以上.培养基MS+TDZ 0.05 mg/L+-NAA 0.02 mg/L适合“圣香”苦瓜子叶不定芽分化,分化率为68.4％;培养基MS+-TDZ 0.03 mg/L+-NAA 0.02 mg/L适合“玉华”苦瓜子叶不定芽分化,分化率为67.8％.“圣香”苦瓜在培养基上MS+ZT 0.3 mg/L+NAA 0.01 mg/L上丛生芽诱导效果好,增殖率达6.5;“玉华”苦瓜丛生芽诱导的最佳培养基为MS+TDZ 0.02 mg/L+NAA0.01 mg/L,增殖率达6.6.生根诱导以1/2MS+NAA 0.05 mg/L培养基诱导率最高,“圣香”苦瓜生根率为90.7％,“玉华”苦瓜生根率达91.9％.     

Examining strain propagation in the branch unions of Quercus alba L. and Quercus montana Willd.

Branch failure resulting from static loading events can be caused by ice, snow, or a climbing arborist aloft in a tree and can cause power outages, road blockages, costly property damage, or personal injury. In previous studies, researchers calculated bending stress to investigate the load-bearing capacity of branches. Yet, these calculations require geometric measurement of the wood that may not always be readily measure post failure. As stress and strain are linearly related, this study used static loading trials and digital image correlation to measure strain, tissue displacement, to gain insight on how loads move through the branch union zone. The results indicate that strain is located distally in the branches and diminishes through the branch union zone, suggesting that branch unions may serve a role in the damping of loads in trees.     

Adventitious root formation in Anacardium occidentale L. in response to phytohormones and removal of roots

Despite advances in tissue culture techniques, propagation by leafy, softwood cuttings is the preferred, practical system for vegetative reproduction of many tree and shrub species. Species are frequently defined as ‘difficult’- or ‘easy-to-root’ when propagated by conventional cuttings. Speed of rooting is often linked with ease of propagation, and slow-to-root species may be ‘difficult’ precisely because tissues deteriorate prior to the formation of adventitious roots. Even when roots form, limited development of these may impair the establishment of a cutting.     

Embryo germination and proliferation in vitro of Juglans regia L.

This paper presents the optimal culture conditions for the in vitro embryo germination and proliferation of Juglans regia L. rootstock cv. Peralta, selected by the IMIDA fruticulture team. J. regia L. rootstock cv. Peralta is characterised by its resistance to salinity, lime and by its vigour. The first experiment determined the best culture medium for in vitro embryo germination. The Murashige and Skoog medium (MS; Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissues cultures. Physiol. Plant. 15, 473–497), the medium developed by our team for walnut (NGE), the Lloyd and McCown medium (WPM; Lloyd, G., McCown, B., 1981. Commercially feasible micropropagation of mountain laurel, Kalmia latifolia, by the use of shoot tip culture. Proc. Plant Prop. Soc. 30, 421–427) and that of Driver and Kuniyuki (DKW; Driver, J.A., Kuniyuki, A.M., 1984. In vitro propagation of Paradox walnut rootstock. HortScience 19 (4), 507–509), were compared, all without growth regulators. The best germination percentage was obtained in WPM (81%) with significant differences between the different media. In the second experiment, the optimal benzylaminopurine and indole butyric acid concentrations were determined for the proliferation stage of the explants obtained in the first experiment. The proliferation rates obtained varied from 0 in the medium without cytokinins to 6 in the medium with 0.5 mg l⁻¹ BAP. The cluster proliferation quality and other parameters studied indicate that the optimal treatment was 0.5 mg l⁻¹ BAP.     

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.     

Shoot proliferation and rooting in vitro of Theobroma cacao L. type Amelonado

Cocoa shoot tip and nodal bud expiants were cultured on modified Murashigè and Skoog (1962) media. Shoot proliferation occurred during the first six weeks on media containing BAP, zeatin or zeatin riboside. The shoots so produced were rooted on a medium with IBA, NAA and phloroglucinol.