甘蓝型油菜下胚轴和带柄子叶再生体系研究

以甘蓝型油菜野油19号的下胚轴和带柄子叶为外植体,研究其下胚轴和带柄子叶在不同浓度激素配比下的分化率及再生频率的变化。该品系的油菜下胚轴和带柄子叶在1.5 mg/L的2,4-D中预培养4 d后,转入分化培养基中,其愈伤组织形成早,发生快,再生频率和分化频率均较高。其中,下胚轴转入MS+3 mg/L 6-BA+0.1 mg/L NAA培养基中的分化率和再生频率最高,再生频率达到86.67%;带柄子叶外植体的再生频率要比下胚轴的再生频率低,在MS+4 mg/L 6-BA+0.3 mg/L NAA的分化培养基中芽的再生频率为46.67%。本研究初步建立了甘蓝型油菜野油19号的高频率再生体系。     

甘蓝型油菜子叶和下胚轴再生植株无性系建立

以甘蓝型油菜(Brassica napus L.)豫油2号和6257的子叶和下胚轴为材料,在不同激素配比的MS培养基上诱导出了愈伤组织。将经过继代的部分愈伤转入分化培养基,分化结果表明:除基因型、外植体和分化培养基的激素配比对分化率有影响外,诱导愈伤培养基的激素配比对分化率也至关重要。豫油2号的子叶和下胚轴在最适诱导培养基(ZT 1+NAA0.5+2,4-D 0.2 mg/L)和最适分化培养基(ZT4+IAA 0.2 mg/L)组合中的愈伤分化率分别为12.5%和75%;6257的子叶和下胚轴在其最适诱导培养基(KT 2+NAA1+2,4-D 0.2 mg/L)和最适分化培养基(6-BA 4+IAA 0.02 mg/L)组合中的愈伤分化率分别为50%和37.5%。将其最适诱导培养基中的愈伤组织继代达8个月以上,建立了不同继代愈伤的再生植株无性系。     

白桦下胚轴再生系统的建立

本文对白桦下胚轴的再生系统作了研究。讨论了不同的基本培养基、激素组成、培养条件及外植体的生理年龄在不同阶段产生的效应。通过诱导、分化和生根阶段, 得到了再生植株, 提出了最适的基本培养基、激素组成、培养条件及下胚轴外植体的生理年龄, 为白桦的遗传转化奠定了基础。     

二斑叶螨对不同品种茄子的选择性与适合度

【目的】为明确二斑叶螨Tetranychus urticae Koch对不同品种茄子的选择性与适合度。【方法】通过二斑叶螨对不同品种茄子的田间选择性试验以及在室内不同品种茄子对二斑叶螨种群数量、个体生长发育和繁殖的影响试验,分析了二斑叶螨对9个茄子栽培品种和1个野生茄子品种的寄主选择性,以及在不同品种上的种群动态、发育历期和繁殖力。【结果】二斑叶螨对野生品种蒜芥茄Solanum sisymbriifolium的选择性最差,在该品种上除卵可以正常孵化外其他时期均不能存活。在茄子S.melongena品种间,二斑叶螨对京茄6号和日友长直壮青长茄的选择性最强,分别达到195和183头/株,对布利塔和长竹丝茄的选择性最差,分别为63.5和59.75头/株。接入20对成螨后第28天,紫龙王4号上的数量最多(1 009头/株),京茄6号和嫁接的京茄6号上的数量次之(分别为981和909头/株)。二斑叶螨在京茄6号上的发育历期最短,为9.31 d;在长丰2号上发育历期最长,为11.08 d。京茄6号、京茄黑霸和嫁接的京茄6号上二斑叶螨开始产卵的前5 d内单雌产卵量分别为35.92,33.20和31.34粒/头,是产卵量较多的3个品种,且3个品种之间差异不显著;紫龙王4号、茄砧1号、布利塔和长丰2号上的产卵量较少,分别为18.56,19.24,22.26和23.36粒/头,且4个品种之间差异不显著。【结论】二斑叶螨对紫黑圆茄类的京茄6号的选择性较强、适合度较高,对紫黑长茄布利塔的选择性和适合度为中等,对野生品种蒜芥茄的选择性和适合度表现为最差。     

提高榨菜离体培养植株再生频率

采用榨菜“浙桐1号”品种为材料,以MS为基本培养基,通过对不同植物生长调节剂的组合和不同外植体等主要因素的筛选,大幅度提高了榨菜离体培养植株再生频率。结果表明,2mg／L6．BA 0．2mg／L2,4-D的组合较为适宜,其不定芽再生频率可达50％,且外植体以下胚轴为好：而CPPU和2,4-D的适宜组合为1．5mg／L 0．2mg／L,其不定芽再生频率高达66．67％,最适外植体为带柄子叶。同时,研究结果显示,添加0．25～1mg／L的GA,对榨菜已分化的不定芽的伸长有抑制作用;子叶柄和下胚轴外植体的分化具有极性现象。     

大白菜下胚轴离体不定芽高效再生体系的研究

以大白菜的下胚轴为外植体,比较了不同浓度TDZ和6-BA两种细胞分裂素与不同浓度NAA相配合的培养基上不定芽再生的差异,并利用筛选出的高效再生培养基研究外植体苗龄、切段来源、接种方式以及品种对不定芽再生的影响。结果表明:与6-BA相比,TDZ对诱导下胚轴不定芽再生更有效,在M S+TDZ 0.3 m g.L-1+NAA0.5 m g.L-1+A gNO35 m g.L-1的培养基上,下胚轴不定芽再生频率高达87.8%,平均每下胚轴再生不定芽数也达到15.1个;3~5 d苗龄之间的下胚轴不定芽再生能力无显著差异,再生频率均达到80%以上,此后随着苗龄的增加,不定芽分化频率快速下降,苗龄为7 d时再生频率只有51.1%;下胚轴不同切段不定芽再生能力由强到弱表现为:上部切段>中部切段>下部切段;以正插(形态学下端插入培养基)方式接种的外植体不定芽再生能力显著大于反插(形态学上端插入培养基)和平放的;不同品种大白菜下胚轴的不定芽再生能力有一定差异。     

辣椒离体培养及再生体系的研究

选用9个辣椒(Capsicum annuumL.)品种(系),研究了不同激素组合、基因型、外植体类型、苗龄和Ag-NO3等因素对外植体不定芽分化和伸长的影响.结果表明,在6-BA/IAA为10∶1配比下,有利于辣椒外植体的分化再生,而6-BA/IAA为3∶1配比下适合于再生芽的伸长;不同品种辣椒的再生能力差别较大,分化率在13.3%~90.0%之间;辣椒子叶再生能力比下胚轴强,是较好的外植体材料;12~16 d苗龄的外植体分化频率较高;添加4mg?L-1AgNO3可使芽分化率平均提高16.9%.通过比较,筛选出了适合于辣椒芽分化的培养基为MB5(MS无机盐 B5有机成分) 5 mg?L-16-BA 0.5 mg?L-1IAA 4 mg?L-1AgNO3,芽伸长培养基为MB5 3 mg?L-16-BA 1 mg?L-1IAA 2 mg?L-1GA3 4 mg?L-1AgNO3,生根培养基为1/2 MS 0.2 mg?L-1IAA 0.1 mg?L-1NAA.     

甜椒的离体培养及建立再生体系研究

以荷兰进口甜椒的子叶和下胚轴为外植体,接种到附加不同植物生长调节剂的培养基上,筛选出MS+6-BA 4.0 mg/L+NAA 0.1 mg/L为子叶和下胚轴最佳不定芽分化培养基;MS+IBA 0.2 mg/L和MS+IBA 0.5 mg/L为最佳不定根分化培养基。     

两种茄子子叶诱导胚状体和植株再生

植物名称:茄子(Solanum melongena)品种:远景茄,荷兰茄。材料类别:子叶、下胚轴。培养条件:(1)种子培养基:1/2 SH+1％蔗糖,(2)发生愈伤组织的培养基:MS+2,4-D1mg/L(单位下同),(3)愈伤组织分化培养基:MS附加各种激素,(4)胚状体苗培养基:无激素的MS培养基。培养温度25±2℃,光照强度1200～1400lx,每天照光10～12小时。生长与分化情况:1.愈伤组织的诱导:经常规消毒的种子播于培     

高温胁迫下不同茄子品种幼苗耐热性研究

以14个茄子品种为材料,研究了高温对茄子幼苗的热害指数、恢复指数、电解质渗透率、脯氨酸、可溶性糖含量的影响。结果表明,43／38℃(昼／夜)高温处理下,茄子幼苗热害指数升高,电解质渗透率和脯氨酸含量增加,可溶性糖含量下降。其中。热害指数、恢复指数、电解质渗透率、脯氨酸含量可作为不同茄子品种耐热性的筛选指标。而可溶性糖含量不宜作为耐热筛选指标。高温胁迫条件下,N1(农友茄)表现出极强的耐热性。C2(刺红茄)、F3(丰研二号)、F4(伏龙茄)、Y5(圆杂二号)、X6(西安绿茄)、B7(北京六叶)、Y8(野茄2号)、N9(粘毛茄)、B11(北京五叶)耐热性中等,K11(快圆茄)、R12(二苠茄)、C13(赤茄)、G14(刚果红茄)耐热性极差。     

Influence of silver nitrate (ethylene inhibitor) on cucumber in vitro shoot regeneration

The effect of addition of silver nitrate (AgNO₃) on organogenesis of proximal and distal cotyledon and hypocotyl explants of five cucumber (Cucumis sativus L.) cultivars was investigated. Distal cotyledon and hypocotyl were unresponsive while only poor shoot regeneration was observed in proximal cotyledon and hypocotyl explants of all cucumber cultivars. The addition of different concentrations of AgNO₃ (10, 30 and 50 μM) to the medium, however, induced shoot regeneration in distal cotyledon except Suyo Long cultivar and effectively increased shoot regeneration response as well as the number of shoots per explant in proximal cotyledon and hypocotyl of all cucumber cultivars. This revised version was published online in June 2006 with corrections to the Cover Date.     

Genotype, explant and position effects on organogenesis and somatic embryogenesis in eggplant (Solanum melongena L.)

The relative importance of genotype and explant, and their interactionsfor in vitro plant regeneration via both organogenesis and somaticembryogenesis in Solanum melongena (eggplant) has been studied.Hypocotyl, cotyledon and leaf explants of four commerciallygrown Indian cultivars, Pusa Purple Long, Long White Cluster,Pusa Kranti, and Pusa Purple Cluster were used in the study.A combination of benzyladen-ine (11.1 µM) and indoleaceticacid (2.9 µM) was found to be optimum for shoot regeneration.Naphthalene acetic acid induced embryogenesis in all the threeexplants; 32.2µM was optimum for hypocotyl explants while10.7µM yielded maximum number of somatic embryos fromcotyledon and leaf explants. Genotype, explant and genotype-explantinteraction had highly significant effects on both organogenesisand somatic embryogenesis with genotype exerting maximum effecton both these processes. Pusa Purple Long was found to be themost responsive genotype for regeneration of both adventitiousshoots and somatic embryos among the cultivars. Among the explants,hypocotyls yielded the maximum number of adventitious shootsfollowed by cotyledons and leaves. The embryogenic responseof leaves and cotyledons was, however, significantly higherthan that of hypocotyl explants. Significant differences formorphogenetic potential were also observed within a single explant(hypocotyl). There was a basipetal gradient for organogenesis(i.e. decrease in number of shoots from base to apex) whilethe terminal hypocotyl segments showed better embryogenic potentialthan the median segments. Key words: Solarium melongena, organogenesis, somatic embryogenesis, genotype, explant, position effect     

The optimization of regeneration tissue culture system of three chilli peppers cultivars based on the uniform design and the mathematical model equation

Using uniform random design optimization and the mathematical model equation we optimized the regeneration tissue culture system of the chilli pepper. An efficient and detailed plant reproducible protocol in vitro has been established using different explants and induction media for three chilli pepper cultivars. The result displayed that the seedlings at the curved hypocotyl stage were the best choice to prepare for explants, the genotype of explants affected shoot buds induction frequency and number of shoot buds per explant, and the cotyledon explant was more responsive than hypocotyl explant. The optimal media for maximum shoot initiation and regeneration and the optimal elongation medium were obtained. For Capsicum annuum var. annuum (cv. Xinsu), Capsicum annuum var. annuum (cv. Neimengchifeng) and Capsicum frutescens (cv. Xingfu), the induction rates were 99.17%, 97.50 and 96.11%, respectively; the elongation rates of shoot buds were 86.67%, 85.19% and 82.96%, respectively. The MS medium with 0.57 μM IAA and 0.69 μM NAA is the best choice for root induction. The frequency of their root emergence was 95.00-98.33%. Regenerated chilli peppers were successfully acclimatized and cultivated with 100% survival. This work will help to improve multiplication process and the genotype of chilli pepper, and may have commercial impact.     

Factors influencing plant regeneration from seedling explants of Hairy nightshade (<Emphasis Type="Italic">Solanum sarrachoides</Emphasis>)

Hairy nightshade (Solanum sarrachoides) has the potential to be a model system for the study of plant-pathogen interactions, however, the availability of tissue culture and transformation methods would strengthen its utility. For the development of tissue culture methods, we investigated, explant type (cotyledons, hypocotyls, roots), hypocotyl explant origin, cotyledon orientation (abaxial vs. adaxial) in direct contact with the medium, gelling agents (agar and agargel) and cytokinins (zeatin and 6-benzyladenine) at different concentrations. Cotyledon explants resulted in the greatest biomass as compared to root and hypocotyl. As for hypocotyl explant origin, explants proximal to the cotyledons had a significant effect on plant regeneration. However, cotyledon orientation and gelling agent had no effect on plant regeneration. Medium supplemented with either zeatin or 6-benzyladenine at 1 mg L⁻¹ resulted in significant shoot regeneration. Shoots rooted readily when cultured on a non-hormone based rooting medium.     

Correlated response of in vitro regeneration capacity from different source of explants inCucumis melo

The variation among and within different populations of the regeneration ability from leaf, cotyledon and hypocotyl explants has been studied. A control population and two lines selected by their regeneration capacity from leaf explants were used. Significant differences among the plants of the control population,for the organogenic response, were detected. The regeneration capacity varies depending on the type of explant. Selection in order to improve the regeneration frequency from leaf explants also raises the organogenic response in the other explant types. This result suggests the presence of a partial common genetic system controlling the regeneration frequency of the diverse types of explants.Abbreviations BA 6-benzylaminopurine - IAA indole-3-acetic-acid - K 6-furfuryl-aminopurine - NAA 1-naphthalene acetic acid     

In vitro regeneration of sesame (Sesamum indicum L.) from seedling cotyledon and hypocotyl explants

The goal of this study was to develop an efficient regeneration protocol to be used for genetic transformation of sesame. Published regeneration methods using benzyladenine (BA) and 1-naphthalene acetic acid (NAA) were unsuccessful for the cultivars used herein. Experiments were carried out using cotyledon and hypocotyl explants from the cultivar Mtwara-2. Later the optimised culture conditions were used to investigate the regeneration response of different genotypes. There was significant interaction between hormone treatments and macronutrients for shoot and root regeneration. Results also showed that shoot regeneration was significantly influenced by explant type. Shoots were only obtained from cotyledons whereas both cotyledons and hypocotyls could produce roots. Modified Murashige and Skoog (MS) medium with N6 macronutrients resulted in twice the shoot regeneration frequency obtained with ½MS macronutrients in the presence of thidiazuron (TDZ). The shoot regeneration frequency was significantly reduced when BA was used in place of TDZ. On shoot regeneration medium containing BA and NAA, only roots were formed. Replacing NAA with indole-3-acetic acid (IAA) greatly improved the regeneration of shoots. The optimum growth regulator combination for shoot regeneration was 20 μM TDZ together with 2.5 μM IAA, which gave a frequency of 63% and 4.4 shoots per regenerating explant for the best cultivar Ex-El. Genotypic differences were significant both for the number of explants regenerating shoots and the number of shoots produced per regenerating explant.     

Thidiazuron-induced high-frequency axillary and adventitious shoot regeneration in Vigna radiata (L.) Wilczek

Summary Prolific shoot regeneration was achieved in mungbean Vigna radiata (L.) Wilczek from 3-d-old in vitro cotyledonary node and hypocotyl explants from seedlings derived from mature seeds on Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ) (0.9 μM). An initial exposure to TDZ for 20 d and three successive transfers to fresh medium with reduced thidiazuron levels (0.09 μM) resulted in the regeneration of 104 shoots/explant from the cotyledon and 30 shoots/explant from the hypocotyl. Thidiazuron-associated abnormalities such as short compact shoots, fasciation and leaf growth in the form of rosettes were observed in shoots regenerated from hypocotyl explants. Both axillary and adventitious shoot formation from the explants were confirmed by histology. Through repectitive cycles of regeneration in the presence of TDZ, the number of shoots that could be obtained from the two explant classes within 80 d was significantly higher than with previous reports in mungbean     

The Effect of a Submersion Pretreatment on In Vitro Explant Growth and Shoot Regeneration from Hypocotyls of Flax (Linum Usitatissimum)

This study was carried out to determine the effect of temporary submersion of hypocotyl segments in water on in vitro explant growth and shoot regeneration on MS (Murashige and Skoog, 1962) medium supplemented with 1 mg l⁻¹ BAP (6-benzylaminopurine) and 0.02 mg l⁻¹ NAA (naphthaleneacetic acid) in three flax cultivars. It was observed that water-treated hypocotyl explants gave rise to the highest values with respect to shoot regeneration percentage, shoot number per hypocotyl, shoot length and total shoot number per Petri dish, successful rooting and plantlet establishment. This procedure may be applicable for other species cultured in vitro.     

High Frequency of Shoot Regeneration from Hypocotyls and Stem Segments of Antirrhinum majus (Snapdragon)

A broadly applicable direct shoot regeneration method from hypocotyls and stem explants has been developed for six cultivars of Antirrhinum majus L. In order to establish a stable and high frequency of shoot regeneration system, leaves, hypocotyls and stem explants of six cultivars were tested with 72 combinations of auxin (naphthaleneacetic acid (NAA) or 3-indoleacetic acid (IAA)) and cytokinin (6-benzylaminopurine (BA) or zeatin (Z)). A few adventitious shoots were directly regenerated from hypocotyl segments of cv. Orchid on MS medium with NAA + BA, IAA + BA, NAA + Z and IAA + Z. High frequency of direct shoot regeneration was obtained from hypocotyl segments on MS medium with 0.05, 0.1 or 0.25 mg l⁻¹ NAA + 2 mg l⁻¹ Z and 0.5 mg l⁻¹ IAA + 2 mg l⁻¹ Z. Finally, stable and high frequency (92–100%) of shoot regeneration with more than 10 adventitious shoots per explant was achieved from the hypocotyls and stem explants of all six cultivars on MS medium with 0.25 mg l⁻¹ NAA + 2 mg l⁻¹ Z. The shoots emerged directly from the hypocotyls and stem segments 4 weeks after culture initiation.     

Highly efficient Agrobacterium-mediated transformation and plant regeneration system for genome engineering in tomato

Tomato (Solanum lycopersicum L.) is an important vegetable and nutritious crop plant worldwide. They are rich sources of several indispensable compounds such as lycopene, minerals, vitamins, carotenoids, essential amino acids, and bioactive polyphenols. Plant regeneration and Agrobacterium-mediated genetic transformation system from different explants in various genotypes of tomato are necessary for genetic improvement. Among diverse plant growth regulator (PGR) combinations and concentrations tested, Zeatin (ZEA) at 2.0 mg l^?1 in combination with 0.1 mg l^?1 indole-3-acetic acid (IAA) generated the most shoots/explant from the cotyledon of Arka Vikas (36.48 shoots/explant) and PED (24.68 shoots/explant), respectively. The hypocotyl explant produced 28.76 shoots/explant in Arka Vikas and 19.44 shoots/explant in PED. In contrast, leaf explant induced 23.54 shoots/explant in Arka Vikas and 17.64 shoots/explant in PED. The obtained multiple shoot buds from three explant types were elongated on a medium fortified with Gibberellic acid (GA₃) (1.0 mg l^?1), IAA (0.5 mg l^?1), and ZEA (0.5 mg l^?1) in both the cultivars. The rooting was observed on a medium amended with 0.5 mg l^?1 indole 3-butyric acid (IBA). The transformation efficiency was significantly improved by optimizing the pre-culture of explants, co-cultivation duration, bacterial density and infection time, and acetosyringone concentration. The presence of transgenes in the plant genome was validated using different methods like histochemical GUS assay, Polymerase Chain Reaction (PCR), and Southern blotting. The transformation efficiency was 42.8% in PED and 64.6% in Arka Vikas. A highly repeatable plant regeneration protocol was established by manipulating various plant growth regulators (PGRs) in two tomato cultivars (Arka Vikas and PED). The Agrobacterium-mediated transformation method was optimized using different explants like cotyledon, hypocotyl, and leaf of two tomato genotypes. The present study could be favourable to transferring desirable traits and precise genome editing techniques to develop superior tomato genotypes.