诱导山楂子叶产生不定植株的技术研究

以小金星山楂子叶为外植体，研究了光、细胞分裂素、生长素及蔗糖4个因素对子叶再生不定梢的影响。其影响在小顺序为光＞生长素＞细胞分裂素＞蔗糖。认为山楂胚子叶再生不定植株的最佳条件应当是暗处理3周＋BA3～5mg/L NAA0.1～0．2mg/L 蔗糖4％。生根培养基为1／2MS＋IBA0．3mg/L＋蔗糖2％，先暗培养1周，再转到光下培养。     

甜樱桃品种吉列玛叶片再生不定梢的研究

在基本培养基NN69或WPM上附加BA和低浓度的NAA，IAA，IBA作培养基，诱导甜樱桃品种吉列玛试管苗的叶片产生了不定梢。以WPM附加BA 3mg/L和IAA0．3mg/L的处理，不定梢再生率最高为33．3％。     

Effect of leaf excision time and age,BA concentration and dark treatments on in vitro shoot regeneration of M.26 apple rootstock

SUMMARY

The effect of benzyladenine (BA) concentrations both during the last proliferating subculture before regeneration (10-222 and, 444 µM) and during organogenesis (11.1 and 22.2 µM), leaf excision time (15 and 30 d from the beginning of the subculture), leafage and dark treatments, on adventitious shoot regeneration of M.26 apple roostock were evaluated. Leaves excised 30 d after the beginning of the last proliferating subculture and grown wkhout BA in thè medium gave the highest percentages of organogenesis, while the number of "regenerated shoots per leaf did not differ significantly among the different BA x leaf excision time combinations. The highest BA concentration (22.2 µM) in the organogeneticmedium produced thehighest percentage of regenerating leaves, with no differences between the lengths and numbers of shoots per regenerating leaf. The first twMjnfurled apical leaves showed a greater regenerative ability than the third and fourth ones, whereas the lengths and numbers of regenerated shoots per leaf were similar. The highest leaf organogenetic ratejyas observed when darkness was imposed at the begirP-ning of the last proliferating subculture and/or at the beginning of the organogenetic phase, but more regenerated shoots per leaf were obtained with darkness provided at the beginning or at the end of the lastproliferating subculture; shoot lengths were similar in all the dark treatments. The great influence onorganogenesis of all the treatments applied in the last proliferating subculture indicates the importance of this stage inpreparing explants for shoot regeneration and thus the possibility of using inductive factors in this phase.     

甜樱桃砧木吉塞拉(Gisela)叶片再生体系研究

 以甜樱桃矮化砧木‘Gisela 5’和‘Gisela 6’的组培苗为试材，研究建立叶片的离体再生体系。结果表明，Gisela 6的再生能力高于Gisela 5；叶片正面接触培养基比背面接触培养基更有利于分化再生。用新梢顶端1~3节新展开的叶片制备外植体，分别接种在WPM+BA 7 mg/L+IBA 0.3 mg/L和WPM+BA 5 mg/L+IBA 0.5 mg/L的培养基上，黑暗处理15 d，Gisela 5的最高分化频率为75.1%，Gisela 6为100%。再生芽苗在三角瓶中生根容易，移人大田后生长正常。     

In vitro morphogenesis of juvenile Annona cherimola Mill,bud explants

A micropropagation procedure for juvenile cherimoya (Annona clierimola Mill.) is described. Axillary shoot proliferation was obtained after culturing nodal sections with lateral buds in basal medium supplemented with either 0.66 μM BA or 1,36μM zeatin. For root induction shoots were pre-incubated for 7 d in basal medium supplemented with 1 gl^?1 activated charcoal, then cultured for 10 d (7 dark/3 light) on medium with 500 μM IBA, 58.4 mM sucrose, and 200 mg I“1 citric acid. Afterwards, shoots were transferred to the same medium without auxin and with the macroelements at half strength. Using this procedure, 95% of shoots rooted. The survival rate at the end of the acclimatization period was 70%.     

不同激素和黑暗处理对泰山红石榴离体叶片再生的影响

以泰山红石榴为试材,研究了不同激素配比、黑暗处理时间等关键因素对泰山红石榴离体叶片愈伤组织诱导和不定芽再生的影响。结果表明,细胞分裂素是离体叶片再生不定芽所必需的。生长素2,4-D与IBA组合诱导出的愈伤组织生长量大,但不定芽再生率低。黑暗处理时间以2周为宜。适宜的再生培养基为MS+KT1.0mg·L-1+BA2.0mg·L-1+GA30.5mg·L-1,不定芽再生率达57.0%,每片叶再生芽数为3.0个。     

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。     

秋水仙素诱导樱桃矮化砧木'吉塞拉6号'获得六倍体再生植株

 以三倍体樱桃矮化砧木'吉塞拉6号'（Prunus ceransus × P. canescens）的离体叶片为外植体,采用秋水仙素诱导处理再生出六倍体植株。将外植体首先在加有秋水仙素（50 mg. L-1）、生长素（IBA 0.5 mg. L-1）和细胞分裂素（BA 5.0 mg. L-1）的改良WPM液体培养基中培养5 d,再转移到不含秋水仙素（其它成分相同）的固体培养基上继续培养56 d,再生出形态变异明显的新梢。采用流式细胞仪鉴定染色体倍性,确定其为六倍体新梢。六倍体植株与三倍体的'吉塞拉6号'植株形态学上有明显差异。六倍体的试管苗已在大田移栽成活,并已成功高接在甜樱桃大树上。     

悬铃木下胚轴及叶片再生不定芽的影响因素分析

 以悬铃木的下胚轴和离体叶片为材料, 研究了基因型、植物生长调节剂、光照条件、外植体部位等因素对其不定芽分化的影响。结果表明: 1) 6-BA与IBA的浓度比对下胚轴不定芽的分化影响较大,以6-BA ∶IBA小于20 ∶1较为合适; 而叶片的不定芽再生受基因型的影响显著, 在4种供试材料中, PH2的再生能力最强, 但0.1～1.0 mg·L^-1 TDZ不利于其不定芽再生。2) 黑暗培养对下胚轴的不定芽分化有一定的促进作用, 但不利于叶片的不定芽再生, 50～100 lx的弱光有利于叶片的不定芽分化。3) 不同部位的下胚轴切段及叶片的再生能力差异显著, 其中靠近子叶的下胚轴切段和试管苗顶端的2枚叶片再生能力较强。此外, 刻伤叶片的再生效果明显优于叶片切块, 说明悬铃木的叶片与下胚轴的离体培养存在明显的生物全息现象。     

Micropropagation of red raspberry and the influence of phloroglucinol

Micropropagation of 12 raspberry seedling selections and the cultivar ‘Malling Jewel’ has been achieved. A basic culture medium (Linsmaier and Skoog, 1965) supplemented with benzylaminopurine (BAP), 1.0 mg l^?1, and indol-3-yl butyric acid (IBA), 0.1 mg l^?1, was optimal for shoot proliferation. The presence of phloroglucinol (PG) at a concentration of 162 mg l^?1 significantly increased shoot number at all auxin: cytokinin concentrations. Removal of the cytokinin and increasing the concentration of IBA to 1.0 mg l^?1 resulted in adventitious root formation. PG synergistically promoted the number of roots per rooted culture but did not significantly increase the percentage rooting. Viable plants were produced from all genotypes when transplanted to soil.     

The Internal Blackening of Potatoes Caused By Bruising

Summary

Pandorea jasminoides and P. pandorana are successful Australian endemic ornamental species of the family Bignoniaceae. Vigorous growth and spectacular floral display make them popular landscape plants. Application of tissueculture could facilitate Pandorea breeding and improvement programmes, however, there is no report on in vitro regeneration of the genus Pandorea. A protocol for de novo shoot production is described for commercial cultivars of P. pandorana and P. jasminoides. Amenability of explants under in vitro conditions was found to differ between the two species. Direct shoot organogenesis of P. pandorana was obtained using node explants while seedling (epicotyl) explants of P. jasminoides were found to be responsive for shoot regeneration. A maximum of 13 and 15 shoots per explant of P. pandorana and P. jasminoides, respectively, was obtained. Benzyladenine, BA, (8.8.mM) proved to be more effective for shoot regeneration than thidiazuron (TDZ) for P. pandorana while a combination of cytokinin (BA and N-(choloro-4-pyridyl)-N-phynyl urea (CCPU) and auxin is required for P. jasminoides regeneration. However, no significant difference in rooting of in vitro regenerated P. pandorana and P. jasminoides shoots was observed. Regenerated shoots were rooted on a medium containing indolebutyric acid (IBA) and plantlets were successfully established under glasshouse conditions.     

Black Currant Leaf Spot: II. Laboratory Tests of Fungicides for the Prevention of Sporing of Pseudopeziza Ribis on Overwintered Leaves

Summary

We have developed a two-step procedure for rooting of tea microshoots in vitro. The effectiveness of different auxin treatments for root formation was found to differ. Among the auxins tested, 25 μM -naphthalene acetic acid (NAA) gave the best results, with 100% rooting, compared to 25 μM indole-3-butyric acid (IBA) or 25 μM indole-3-acetic acid (IAA), which induced 17% and 58% rooting, respectively. Incubation of tea microshoots on 0.33 Murashige and Skoog (MS) medium, supplemented with 25.0 μM NAA or 175.0 μM IBA for 10 d, followed by transfer to auxin-free 0.33 MS medium resulted in 100% rooting, whereas 50.0 μM IAA induced 91.7% rooting. Besides the different auxin treatments, the strength of the MS medium, the duration of incubation of microshoots in auxin-containing medium, the sucrose concentration, the gelling agent, the pH of the medium, the incubation temperature, the light intensity, and the quality of the shoots also played a significant role during in vitro rooting of micropropagated tea shoots. Among the combinations tested, the most effective results were obtained when green microshoots were incubated on 0.33 MS medium supplemented with 25.0 μM NAA, 50.0 mM sucrose, pH 5.5, gelled with 0.2% (w/v) Phytagel^TM for 10 d at 25° – 30°C at a light intensity of 40 μmol m^–2 s^–1, followed by transfer of shoots to auxin-free 0.3 MS medium. This resulted in 100% rooting and, on average, 11 long roots were formed per shoot. Anatomical changes during adventitious rooting of micropropagated tea shoots in vitro were also studied to understand the process of rooting.     

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。     

香玲核桃离体叶片再生体系的建立

为解决核桃种质资源的实验室保存及转基因研究等问题,研究了暗培养时间、叶片放置方式及不同浓度激素组合(BA、NAA、TDZ)对香玲核桃叶片再生的影响。试验结果表明,最适宜的暗培养时间为14d;近轴面放置效果最好;不同组合的激素配比对叶片再生不定芽能力不同,最适宜香玲核桃叶片再生的培养基为:MS+BA0.5mg·L-1+NAA0.5mg·L-1+TDZ1.0mg·L-1+sucrose30g·L-1+agar6g·L-1,在该培养基上,离体叶片不定芽再生率达60%,每叶片平均再生不定芽数超过1.1个。筛选得到了香玲核桃离体叶片的最适分化及生根培养基,对离体再生培养条件进行优化,为香玲核桃的种质资源保存及遗传转化研究奠定了基础。     

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.     

High-frequency somatic embryo production from unfertilized ovules of grapes

Somatic embryos were produced in large numbers from cultured, unfertilized ovules of Vitis vinifera, cultivars ‘Cabernet Sauvignon’ and ‘Grenache’, and of the hybrid grape ‘Gloryvine’ (V. vinifera × V. rupestris). Callus of nucellar origin was produced by culturing the ovule explants initially in Nitsch medium containing 5 μM 2,4-dichlorophenoxy acetic acid (2,4-D) or 5 μM β-naphthoxyacetic acid (NOA) plus 1 μM benzyladenine (BA), and then in a medium supplemented with NOA (10 μM) plus BA (1 μM). Embryos were produced when callus was transferred to basal medium containing no auxin or cytokinin. Secondary and tertiary embryos were formed by budding of primary somatic embryos. Prolonged culture of ovular callus in NOA (10 μM) plus BA (1 μM) gave rise to cell aggregates which developed into free-floating roots when transferred to basal medium.     

Micropropagation of Prunus scoparia,a Suitable Rootstock for Almond under Drought Conditions

Prunus scoparia is a wild deciduous shrub, usually living on dry calcareous soils of the rocky mountains and has been used as a grafting rootstock for domesticated almonds to provide drought resistance. In the current study, micropropagation ability of P. scoparia was investigated using cytokinin and auxin. Uniform nodal shoot pieces (3–5 cm in length) of seedlings were used as explants. The explants were disinfected with 10% sodium hypochlorite solution. For adventitious shoot induction and proliferation, Murashige and Skoog (MS) media containing 7.00 g/l agar and 30.00 g/l sucrose containing five concentrations of benzyl adenine (BA) (0.00, 0.50, 1.00, 2.00, and 4.00 mg/1) and also containing six concentrations of Thidiazuron (TDZ) (0.00, 0.50, 1.00, 2.00, 5.00, and 7.00 mg/1) were compared. For rooting, in vitro shoots (2–3 cm) were transferred into ½ MS medium supplemented with 30 g/l sucrose, 7.50 g/l agar, and different concentrations of IBA (0.00, 0.25, 0.50, and 1.00 mg/l) and NAA (0.00, 0.25, 0.50, and 1.00 mg/l). Based on the results obtained for shoot proliferation, only 2.00 and 4.00 mg/l BA and 2.00 mg/l TDZ concentrations generated shoots, while other treatments did not show shoot proliferation. Among the three treatments that generated shoots, the best results for shoot number, leaf number, and leaf color quality were observed in media containing 2.00 mg/l TDZ. Based on the results obtained for rooting, the effect of IBA concentrations on the rooting percentage, root number, and root length was significant. Among IBA concentrations, only 0.50 mg/l IBA induced rooting, while there was no rooting in the media containing other IBA concentrations. None of the NAA concentrations showed rooting. In conclusion, MS culture medium supplemented with 2.00 mg/l TDZ and ½ MS culture medium supplemented with 0.50 mg/l IBA are suggested for in vitro shoot proliferation and rooting of P. scoparia, respectively. The results presented herein could be used for in vitro selection and micropropagation of P. scoparia.     

培养条件对酸枣叶片不定梢再生率的影响

以酸枣无菌苗叶片为外植体,选用基本培养基ＮＮ６９和ＷＰＭ,外源激素包括ＴＤＺ、ＩＡＡ、ＩＢＡ和ＮＡＡ,暗培养时间为２、３、４周。采用正交实验设计方法,研究了基本培养基、外源激素及暗培养时间对叶片不定梢再生的影响。结果表明在附加ＴＤＺ１~３ｍｇ/Ｌ的ＷＰＭ和ＮＮ６９上都可诱导不定梢再生,但ＷＰＭ比ＮＮ６９更有效。暗培养时间对不定梢再生也有影响,暗培养３周比２、４周更有利于提高叶片的不定梢再生率。在附加ＴＤＺ１ｍｇ/Ｌ、ＩＡＡ０．１ｍｇ/Ｌ的ＷＰＭ培养基上,暗培养３周后转到光下培养,获得的不定梢再生率达８７．５%。     

Callus induction and plant regeneration in Cardiospermum halicacabum Linn. an important medicinal plant

Cardiospermum halicacabum Linn. is an important medicinal twining herb belonging to the family sapindaceae. A method for rapid micropropagation of C. helicacabum through plant regeneration from leaf and nodal explant derived calli has been developed. The nodal and leaf segments were cultured on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D; 0.5–9 μM) for callus induction. Callus production was highest at 5 μM 2,4-D where 96 and 90% of cultured leaf and nodal cuttings produced callus, respectively. The viable calli were maintained at reduced concentration of 2,4-D (2 μM). These calli were transferred to MS medium supplemented with various concentrations of 6-benzyladenine (BA; 2–10 μM) or kinetin (2–10 μM) alone or in combination with indole 3-acetic acid (IAA; 0.2–1.0 μM) for shoot regeneration. The addition of low concentrations of IAA into BA or kinetin containing medium significantly increased the frequency of shoot regeneration in both nodal cuttings and leaf-derived calli. The highest number of adventitious shoots (28 per callus) formed at 8 μM Kin and 0.5 μM IAA. For rooting of the shoots, half-strength MS medium supplemented with different concentrations of indole 3-acetic acid, indole 3-butyric acid (IBA) and (alpha)-naphthalene acetic acid (NAA) 1–5 μM was tried. The optimal result was observed on half-strength MS medium supplemented with 2.5 μM IBA, on which 91% of the regenerated shoots developed roots with an average of 4.2 roots per shoot within 45 days. The in vitro raised plantlets were acclimatized and transferred to soil with 90% success. This in vitro propagation protocol should be useful for conservation as well as mass propagation of this medicinal plant.     

不同基因型梨叶片离体培养和植株再生

 以‘巴梨’、‘身不知’和‘早酥’梨试管苗叶片为外植体, 对不定芽进行了诱导、增殖和生根。重点探讨了基本培养基、植物生长调节剂配比、AgNO₃ 不同浓度和NH4⁺-N与NO3^--N比例对不定芽再生的影响。结果表明, MS + TDZ 0.5 mg/L + IBA 0.1 mg/L为‘巴梨’叶片不定芽发生的最佳培养基。在QL + TDZ 1.0 mg/L +NAA 0.1 mg/L培养基上, 暗培养3周后转光下培养, ‘身不知’和‘早酥’分别获得89.6%、81.2%的不定芽再生率和3.45、3.73的平均再生芽数; 对‘巴梨’和‘早酥’不定芽再生有效促进的AgNO₃ 浓度范围为0.1～0.5 mg/L, 0.1～4.0 mg/L的AgNO₃、1∶2～7的NH₄⁺-N∶NO ₃^--N和21.50mmol/L的K+对‘身不知’叶片再生均有促进作用, 缺乏NH₄⁺-N不利于不定芽的再生; 转移到MS+BA1.0 mg/L + IBA 0.1 mg/L培养基上能快速增殖; 在MS、1 /4MS + IBA 1.0～2.5 mg/L +蔗糖5～15 g/L +活性炭0.5～1.0 g/L上获得了不同程度的生根苗。