An efficient method for immature embryo rescue and plant regeneration from the Calli of Ziziphus jujuba ‘Lengbaiyu’

Hybrid breeding of Chinese jujube (Ziziphus jujube) is limited by embryo abortion. Improving the success of embryogenic progeny and immature embryo culture techniques could increase the breeding efficiency of new jujube varieties. We optimised the current immature embryo culture techniques by using immature embryos before the globular stage of Chinese jujube variety Lengbaiyu. The optimal growth media were as follows: for 30-day-old immature embryo growth, MS + IBA 0.8 mg·L^?1 + ZT 0.5 mg·L^?1 + GA₃ 7.0 mg·L^?1 + NAA 0.2 mg·L^?1 + LH 0.5 mg·L^?1 + 5% sucrose, with 22.22% embryonic growth rate and 0.41% plantlet formation rate; and for young embryo differentiation, MS + TDZ (0.8 mg·L^?1) + IAA (0.5 mg·L^?1) + sucrose (3%), with 81.12% differentiation rate. Thus, regeneration via callus induction effectively improved the efficiency of embryo rescue from young immature embryos in Lengbaiyu, which might provide new approaches for breeding Chinese jujube.     

Effects of culture conditions on in vitro bulblet induction in the medicinal plant Amana edulis (Miq.) Honda

Amana edulis (Miq.) Honda is an important medicinal plant with a variety of anti-cancer properties, and it is of great importance to improve its reproductive rate through micropropagation technology to meet increasing demand. In the present study, in order to establish a rapid in vitro bulblet propagation protocol for A. edulis, an L₁₆ (4⁵) orthogonal design was used to investigate the effects of sucrose, 6-benzyladenine (6-BA), α-naphthaleneacetic acid (NAA), and macro-elements on bulblet induction. The results show that the sucrose concentration was the crucial factor for A. edulis bulblet initiation, followed by 6-BA and macro-elements, while NAA had the weakest effect. The optimal medium for in vitro bulblet induction was Murashige and Skoog medium supplemented with 0.1 mg·L^?1 6-BA, 0.01 mg·L^?1 NAA, and 100 g·L^?1 or 80 g·L^?1 sucrose (pH 5.8), in which A. edulis shoot clusters (without roots) were cultured at 5(±2)°C for 35 d or 60 d and then incubated at 23(±2)°C for 90 d. The entire cultivation process occurred in the dark. The present study demonstrates that this protocol can be used for the propagation of A. edulis.     

Direct somatic embryogenesis in Epipactis veratrifolia,a temperate terrestrial orchid

Most temperate terrestrial orchids are endangered species. Attempts to produce plantlets from plants of the genus Epipactis by asexual methods have totally failed. This study was conducted using somatic embryogenesis as a rapid vegetative propagation technique for conservation of E. veratrifolia. For these purposes, effects of different types of plant growth regulators (PGRs), different types of explants, light and dark conditions, and the effect of gibberellic acid (GA₃) and Paclobutrazol (PBZ) were investigated on somatic embryogenesis induction. Abscisic acid (ABA) pre-treatment effectiveness on inducting somatic embryogenesis and increasing the number of embryos per explant were investigated. Subsequently, 16 media were tested to find the best medium for plant regeneration and shoot and root proliferation. BA (3 mg L^?1) resulted in a better response than the other PGRs by supporting the development of 17 embryos per protocorm. PBZ, which resulted in 11 embryos per explant, was better than GA₃. FAST medium supplemented with organic substances was recognized as the best medium for plant regeneration and shoot and root proliferation. ABA pre-treatment had positive effect on somatic embryogenesis initiation. This study, for the first time, succeeded in finding a rapid and suitable protocol for propagation and genetic resource conservation of E. veratrifolia.     

山杜英离体培养植株再生的研究

 研究了山杜英[Elaeocarpus sylvestris(Lour．)Poir．]带芽茎段的离体培养及植株再生。筛选出最佳培养基：(1)启动培养：Ms+BA 1.0～2.0 mg·L^-1 (单位下同)+NAA 0.05+蔗糖3%；(2)愈伤组织发生型丛生苗增殖培养：MS+BA 1.0+NAA 0.5+蔗糖3%；腋芽发生型丛生苗增殖培养：MS+BA2.0+IBA 0.1+蔗糖3%；(3)有效苗诱导培养：MS+BA 0.5+IBA 0.1+GA 1.0+蔗糖3%；(4)生根培养：1/2 MS+IBA 3.0+蔗糖2%。用透气膜封FI比用聚乙烯菌膜生根率明显提高，生根明显提前。     

珍稀濒危植物珙桐离体快繁技术初步研究

 研究了我国一级珍稀濒危树种珙桐(Davidia involucrata Baill.) 带芽茎段的离体培养。筛选出最佳培养基: ( 1) 冬芽诱导培养基: WPM+BA 2.0 mg·L^-1+NAA 0.1 mg·L^-1; (2) 丛生芽诱导培养基: WPM + BA 2.0 mg·L^-1; (3) 丛生芽增殖培养基: WPM+ BA 2.0 mg·L^-1+ ZT 0.1 mg·L^-1; (4) 生根培养基: WPM + NAA 0.5 mg·L^-1。     

An improved micropropagation protocol for the recalcitrant plant Capsicum – a study with ten cultivars of Capsicum spp. (C. annuum,C. chinense,and C. frutescens) collected from diverse geographical regions of India and Mexico

Capsicum spp. is a commercially important crop of the Solanaceae family, well-known for its multipurpose use as a vegetable, spice, medicinal and ornamental plants. The genus Capsicum is a recalcitrant species in terms of in vitro morphogenesis and plant regeneration. An efficient method was developed for multiple shoot regeneration in 10 cultivars of Capsicum collected from diverse geographical regions of India and Mexico. Seeds germinated in vitro on a half-strength Murashige and Skoog (MS) medium supplemented with 3.0 % sucrose. Nodes of the in vitro germinated seedlings were used as explant for micropropagation. The combination of the 6-benzylaminopurine, indole-3-acetic acid, and spermidine was found to be the best for multiple shoot induction. However, the optimum responcse varied accompanied by different cultivers with maximum 8.9 ± 0.52 (Capsi-10) to 15.3 ± 0.69 (Capsi-5) multiple shoot per explant. Depending on the cultivar, multiplied shoots were successfully rooted with maximum 18.4 ± 0.20 (highest for Capsi-9) to 36.8 ± 0.29 (highest for Capsi-5) roots per shoot on half-strength MS medium supplemented with 2.0 mg l^?1 indole-3-butyric acid, 1.0 mg l^?1 α-naphthalene acetic acid, and 1.5 mM spermidine. Finally, the micropropagated plantlets were acclimatized with 40.0–86.7 % survival rate, depending on different cultivars.     

梨矮化中间砧S2 、S5 和PDR54的离体培养研究

 以梨矮化中间砧S2、S5和PDR54试管苗为试材, 探讨了培养基和植物生长调节剂对梨矮化中间砧试管苗增殖和生根的效应。其结果表明: MS培养基较QL、AS和WPM培养基更有利于中间砧茎的增殖, 2～3 mg·L ^{- 1} 6-BA、0.1～0.2 mg·L ^{- 1} IBA和1～2 mg·L ^{- 1} GA₃组合能有效促进S2、S5和PDR54试管苗的增殖, 其增殖倍数达3.71～5.83。综合各品种增殖倍数、茎高度以及茎质量等指标表明, MS + 3.0 mg·L ^-1 6-BA + 0.2 mg·L^-1 IBA + 2.0 mg·L^-1 GA₃是S2的最佳增殖培养基; MS + 2.0 mg·L^-1 6-BA + 0.1 mg·L^-1 IBA + 1.0 mg·L^-1 GA₃是S5和PDR54增殖的最佳培养基。同时, S2、S5和PDR54增殖的试管苗在不同的生根条件下分别获得了67%、50%和86%的生根率。     

车轮梅茎段高效再生体系的建立

 以车轮梅 (Rhaphiolepis indica L.)茎段为外植体。探讨基本培养基种类、植物生长调节剂组合、蔗糖浓度和AgNO3等因素对茎段器官发生和植株再生的影响。结果表明：MS+6-BA 2.0 mg·L^-1 +NAA 0.5 mg·L^-1 + AgNO3 1.0 mg·L^-1+蔗糖40 g·L^-1培养基最适合不定芽的分化和增殖,不定芽分化率达90%以上,平均每外植体分化不定芽数达5.38个。不定芽可在1/2MS培养基中有效伸长,适宜生根培养基为1/2MS+NAA 1.0 mg·L^-1,生根率达到100%。     

PLBs induction and clonal plantlet regeneration from leaf segment of commercial hybrids of Phalaenopsis

Induction of Protocorm-like bodies (PLBs) from leaf segments is a clonally propagation technology that used somatic embryogenesis regeneration from in vitro buds of floral stalks, and is a potential alternative for replacement actual low-yield and high-cost clonal propagation of Phalaenopsis orchids. The objective of this work was to evaluate induction of PLBs in leaf segments of two commercial hybrids of Phalaenopsis, arranged in culture media with different combinations of plant growth regulators. The leaf segments (0.4–0.5 cm²) used for PLBs inductions were obtained from young in vitro-induced shoots from inflorescence stalks. They are cultivated in NDM culture media, supplemented with combinations of plant growth regulators Naphthaleneacetic Acid, Thidiazuron and Benzyladenine. The flasks were kept for 60-d in darkness and then for cold white light with photon flux density of 35–40 μmol m^?2 s^?1. The cultivar ‘Ph908’ showed a higher percentage of leaf segments with PLBs (45%) and number of PLBs (25 per leaf segment), in medium containing 0.25 mg L^?1 of TDZ, whereas ‘RP3’ showed only 10% containing PLBs and 2 PLBs per leaf segment in NDM with 1.0 mg L^?1 NAA, 20 mg L^?1 BA and 0.125 mg L^?1 TDZ.     

芜菁高频率再生体系的建立及优化

以3个品种芜菁的带柄子叶和胚轴为外植体,采用苯基噻二唑基脲（thidiazuron,TDZ）配合NAA以及BA配合NAA两种组合研究了适于芜菁离体再生的外植体类型和激素组合。发现带柄子叶为外植体,TDZ 7.0 mg·L^-1 + NAA 1.0 mg·L^-1的组合对不定芽分化最为有利。以此离体再生体系为基础,针对AgNO3浓度、苗龄、2,4-D预培养时间3个因素进行优化,得到了芜菁高频率离体再生体系。结果表明：采用5 d苗龄的带柄子叶作为外植体,在含有2,4-D 1.0 mg·L^-1的MS培养基上预培养2 d,添加TDZ 7.0 mg·L^-1 + NAA 1.0 mg·L^-1 + AgNO3 5.0 mg·L^-1的MS培养基为分化培养基对芜菁离体再生最为有利。依品种不同,最高分化率可以达到90%左右。分化后得到的不定芽在IBA 0.1 mg·L^-1的MS培养基上可以100%生根,移植成活率达95%。     

同源四倍体青花菜离体再生及其倍性鉴定

以同源四倍体青花菜带柄子叶为外植体,研究其离体再生体系。结果表明： 10 d苗龄外植体在MS + 0.05 mg·L^-1 NAA + 3.0 mg·L^-1 6-BA+0.8% 琼脂 + 3% 蔗糖培养基上不定芽的再生频率最高为63.3％;继代培养(MS + 0.04 mg·L^-1 NAA + 4.0 mg·L^-1 6-BA + 0.8% 琼脂 + 3% 蔗糖)增殖系数为6,诱导(1/2MS + 0.1 mg·L^-1 NAA + 0.7% 琼脂 + 3% 蔗糖)生根率100％,移栽成活率90％。流式细胞技术分析及染色体计数鉴定结果表明,再生植株的DNA相对含量为二倍体的2 倍,染色体数为2n=4x=36。     

连香树离体快繁初步研究

 连香树为我国珍稀濒危树种, 具有较高的经济价值和观赏价值。本文研究了3年生连香树(Cercidiphyllum japonicum Sieb. et Zucc. ) 带芽茎段的离体培养。筛选出最佳培养基: (1) 腋芽诱导培养基: MS +NAA 0.01 mg·L ^{- 1} ; (2) 丛生芽诱导培养基: MS +BA 1.0 mg·L ^{- 1} ; (3) 丛生芽增殖培养基:MS +BA 2.0 mg·L ^{- 1} + 2,4-D 0.01 mg·L ^{- 1} ; (4) 生根培养基: 1 /2MS + IBA 1.0 mg·L ^{- 1}。     

无核葡萄与中国野生葡萄杂种的胚挽救技术研究

 通过无核葡萄与中国野生葡萄杂交, 获得了4个杂交组合的后代植株, 确定了各杂交组合取胚珠培养的最佳时期。1Flame ×山葡萄; 2红宝石×爱莫无核; 3红宝石×北醇; 4爱莫无核×山葡萄在授粉后45 d取样培养效果较佳; 5长穗无核白×山葡萄授粉后30 d; 6无核白自交在授粉后35 d培养效果较佳。以NitSchs为基本培养基, 附加0.5 mg·L ^{- 1} 6-BA + 0.5 mg·L^-1 GA₃ + 2.5 mg·L ^{- 1} IBA + 0.1 mg·L^-1ZT, 适合于1、3、5号杂交组合; 而0.5 mg·L^-1 6-BA + 0.5 mg·L^-1 GA₃ + 2.0 mg·L^-1 IBA + 0.1 mg·L^-1ZT适合2、4、6号杂交组合; 胚萌发培养基为2.0 mg·L^-1 IBA + 1.0 mg·L^-1 6-BA +0.2 mg·L^-1 GA₃ , 适合1、2、3、5、6号杂交组合, 而爱莫×山葡萄在1.5 mg·L ^{- 1} IBA + 1.0 mg·L^-1 6-BA + 0.2 mg·L^-1 GA₃培养基上表现较佳, 生根培养基为1 /2MS基本培养基添加0.15 mg·L^-1 IBA + 0.02 mg·L^-1 6-BA, 它适合1号与5号组合, 0.10 mg·L^-1 IBA + 0.02 mg·L^-1 6-BA适合4号与6号组合。     

不同培养条件对‘丰香’草莓离体叶片再生的影响

 以草莓品种‘丰香’离体叶片为外植体, 探讨了基本培养基、不同细胞分裂素、暗培养、硝酸银浓度以及不同植物生长调节剂组合对不定芽再生的影响。结果表明, 基本培养基中以MS 最为适合,WPM、QL 、AS 培养基均不利于不定芽的再生, 而TDZ 的诱导效果好于BA。以MS 基本培养基附加TDZ2.0 mg·L - 1和IBA 0.8 mg·L ^-1可以使‘丰香’叶片不定芽的再生率高达72.33 % , 平均每叶再生芽5.59个。暗培养14 d 可以将‘丰香’叶片的不定芽再生率提高到90.09 %。硝酸银对于提高‘丰香’叶片的不定芽再生没有明显效果, 但在一定程度上改变了细胞分化的方向。     

Production of genetically uniform plants from shoot tips of Aconitum heterophyllum Wall. – a critically endangered medicinal herb

We report the successful micropropagation of a critically endangered medicinal plant Aconitum heterophyllum Wall., using low concentrations of plant growth regulators (PGRs) and molecular validation of the clonal stocks. The maximum rate of in vitro shoot multiplication was obtained on 1.0 × Murashige and Skoog (MS) medium containing 0.25 mg L^?1 Kinetin (Kn) plus 0.25 mg L^?1 Indole acetic acid (IAA). Up to 100% rooting was obtained 15 for shoots cultured on 1.0 × MS medium supplemented with 1.0 mg L^?1 IAA. Adding 0.25 mg L^?1 2,4-dichlorophenoxyacetic acid (2, 4-D) to 1.0 × MS medium resulted in 100% callus formation, while adding 0.25 mg L^?1 IAA plus 0.25 mg L^?1 Kn to 1.0 × MS medium containing 0.25 mg L^?1 2,4-D resulted in 100% generation of embryogenic callus. Inter-simple sequence repeat (ISSR) marker analysis was carried out to check for possible somaclonal variation in the plantlets obtained after three consecutive sub-cultures. Of the 15 ISSR primers used, 10 were found to be monomorphic, with 95–98% similarity, and were used for cluster analysis by the unweighted pair group using arithmetic averages (UPGMA) method. The results revealed that in vitro-regenerated plantlets did not exhibit any genetic polymorphism.     

云南野生早花象牙参叶片再生体系的建立

 以云南野生的早花象牙参叶片为外植体, 探讨了叶片的不同部位、植物生长调节剂组合、暗培养、水解酪蛋白和椰子汁对愈伤组织诱导和不定芽再生的影响。结果表明: 叶片基部, 暗培养, 水解酪蛋白和椰子汁均有利于愈伤组织的诱导; 暗培养促进了不定芽的再生。适宜早花象牙参叶片愈伤组织形成的诱导培养基为MS + 6-BA 1.5 mg·L ^{- 1} +NAA 0.1 mg·L ^{- 1} +水解酪蛋白800 mg·L ^{- 1} +椰子汁50 mL ·L ^{- 1} , 再生培养基为MS + 6-BA 1.5 mg·L ^{- 1} +NAA 0.1 mg·L ^{- 1} , 增殖培养基为MS + 6-BA 0.6 mg·L ^{- 1} + NAA 0.1 mg·L ^{- 1} , 生根培养基为1/2MS +NAA 0.8 mg·L ^{- 1}。     

The influence of exogenously applied 2,4-D and NAA on fruit drop reduction in pummelo cv. Thong Dee

ABSTRACT

Effect of 2,4-D and naphthalene acetic acid (NAA) on fruit drop reduction in pummelo cv. Thong Dee was investigated in the pummelo growing areas of Nakhon Pathom province, Thailand. Five similar sized and aged of pummelo trees were selected to set up the experiment. Ten mature branches with the same size from each pummelo tree were randomly selected around the canopy for 2,4-D (20 and 40 mg L^?1), NAA (20 and 40 mg L^?1) application and control. All treatments were applied to selected pummelo branches 2 times at full bloom and 2 months after fruit set. The results showed that 20 mg L^?1 NAA (14.84%) and 40 mg L^?1 NAA (12.26%) gave significantly higher percent of fruit retention at 6 months after fruit set. However, leaf total nonstructural carbohydrate concentration analysis showed that 40 mg L^?1 2,4-D (104.86 mg g^?1) and 20 mg L^?1 2,4-D (96.55 mg g^?1) gave significantly higher total nonstructural carbohydrate than those in control (78.44 mg g^?1). For fruit quality, 40 mg L^?1 2,4-D and 20 mg L^?1 2,4-D gave the highest peel weight with 435.55 and 358.57 g, respectively, and 40 mg L^?1 2,4-D gave the highest peel thickness with 20.25 mm, while 20 mg L^?1 NAA gave statistically higher total soluble solid than those in 20 mg L^?1 2,4-D and 40 mg L^?1 2,4-D. Therefore, 20 mg L^?1 of NAA sprayed 2 times at full bloom and 2 month after fruit set effectively reduced fruit drop and increased percentage of fruit retention in pummelo cv. Thong Dee.     

光叶子花茎段愈伤组织的诱导及其植株再生的研究

 研究了光叶子花(Bougainvillea glabra Choisy) 茎段愈伤组织的诱导及植株再生。结果表明:愈伤组织诱导以MS +BA 3.0 mg·L ^-1 + 2,4-D 0.2 mg·L^-1 +NAA 0.1 mg·L^-1培养基最好, 茎段愈伤组织的诱导率和分化率分别达78.2%和25.6%。不定芽的产生有两种类型: 直接从茎段基部诱导产生不定芽和茎段基部形成大块愈伤组织后再从愈伤组织上分化出不定芽。丛生苗诱导以MS +BA 2.0 mg·L^-1 +NAA 0.1 mg·L^-1为培养基, 增殖倍数可达5.4, 当培养基中BA 3.0 mg·L^-1和2,4-D 0.5～1.0 mg·L^-1时再生植株会出现叶片变异现象。MS+ BA 0.2 mg·L^-1 + GA₃ 0.5 mg·L^-1 +NAA 0.1 mg·L^-1培养基对有效苗培养具有较好的效果; 生根诱导以MS +NAA 3.0 mg·L^-1培养基诱导率最高, 为88.3%。     

In vitro Germination of Early Ripening Sweet Cherry Varieties (Prunus avium L.) at Different Fruit Ripening Stages

In vitro embryo culture enabled satisfactory germination of immature seeds produced in crosses from early ripening sweet cherry varieties (Prunus avium L.). Three varieties —‘Rita’, ‘Bigarreau Burlat’ and ‘Carmen’— were crossed with ‘Early Star’ as male parent. Germination rate was affected by the developmental stage of both fruit and embryo. Fruit ripening stage was a critical factor for culture infection rate that increased with maturity. In-ovule embryo culture on Murashige and Skoog medium without hormones improved the embryo size but did not increase the germination rate due to a further increase in infection rate. Ex-ovule embryo culture on Murashige and Skoog medium supplemented with BA 1 mg L^?1, NAA 0.5 mg L^?1, 20 g L^?1sucrose, 10 g L^?1 sorbitol and 6 g L^?1agar during the stratification time increased embryo length. Germination was performed on Brooks and Hough medium at the 22?±?1?°C with 16/8 h light/dark photoperiod. The highest germination rate (75?%) was reached in embryos that were 3?4 mm in length, after 30-days stratification at 4?°C. Embryos in fruits at green-yellow stage that were 3?4 mm long were morpho-physiologically developed to produce bipolar seedlings, without combined application of embryo culture and micropropagation.     

辣椒离体植株再生及其变异的SSR检测

采用6个不同基因型辣椒（Capsicum annuum L.）的子叶、下胚轴、去除生长点并带下胚轴和各切去一半的两片子叶、去除生长点并带下胚轴和半片子叶为外植体,分别培养在附加不同激素和 AgNO₃ 的MB₅和MS培养基上,研究不同基因型、外植体、激素配比和AgNO₃等对外植体不定芽诱导和芽伸长的影响。结果表明,不定芽诱导以MB₅+2.0 mg·L^-1 TDZ+1.0 mg·L^-1 IAA+4 mg·L^-1 AgNO₃培养基最佳,诱导率最高可达100 %。而芽伸长的最佳培养基为MB₅+3.0 mg·L^-1 6-BA+0.5 mg·L^-1 IAA+5.0 mg·L^-1 AgNO₃+2.0 mg·L^-1 GA₃,最高伸长率可达87.5 %。生根培养基为MS+0.2 mg·L^-1 IAA+0.1 mg·L^-1 NAA,生根率均达100 %。不同的外植体类型具有不同的不定芽诱导和芽伸长的能力,以去除生长点并带下胚轴和各切除一半的两片子叶作外植体时的诱导效果最好。用50对SSR引物检测再生植株,其中86株伏地尖的再生植株和对照植株间均没有扩增出差异条带,而茄门再生植株的SSR检测结果有12对引物表现多样性。