Imposed starvation for in vitro maintenance and high regeneration of elite key lime (Citrus aurantifolia)

Preservation of elite key lime (Citrus aurantifolia) genotypes using efficient micropropagation and regeneration protocols is very important for new breeding programmes. The objective of the present study was to develop an efficient protocol for micropropagation and maintenance of elite genotypes of key lime based on slow growth and starvation strategy. To do this, regenerated shoots were starved, and the growth was compromised via avoiding subcultures for 4, 8, 12 and 24 months. The numbers of responsive explants and adventitious shoots per each explant were evaluated in the presence of benzylaminopurine (BAP). The results showed that the 12- and 24-month starved explants treated with 1 mg/l BAP were able to produce four times more shoots than the control. In addition, the influences of horizontal or vertical layering explants over medium were evaluated. The results indicated that all horizontally layered explants significantly produced 25% adventitious shoots more than vertical explants, most likely owing to more contact with the surface medium. Finally, it could be concluded that application of slow growth, starvation strategy and also horizontally layering of the explants could efficiently enhance micropropagation and maintenance of threatened elite key lime genotypes with a low cost.     

Interaction of source-plant age and shoot ringing on rooting of cashew (Anacardium occidentale L.) cuttings

Summary

IBA-treated cuttings rooted only when prepared fom bark-ringed shoots. The response to ringing declined with increasing age of the tree. However, on marcotting, the ringed shoots of four year old trees showed more and earlier rooting.     

Factors affecting the shoot multiplication of Persian walnut (Juglans regia L.)

A number of experiments were conducted to identify suitable procedure for in vitro shoot multiplication of Persian walnut (Juglans regia L.). Three different nutrient media (DKW, MS and WPM) and three different gelling agents (Phytagel, Difco Bacto agar and a mixture of Phytagel and Difco Bacto agar) were studied in the first experiment. Driver and Kuniyuki walnut (DKW) medium solidified with 2.2 g l⁻¹ Phytagel was found optimum. Performance of explants was better on DKW medium than on MS and WPM. The DKW and MS media were not significantly different from each other, but both of them were significantly better than WPM, which was a very poor medium for this species. Phytagel alone was significantly better than Difco Bacto agar or Phytagel combined with Difco Bacto agar. In another experiment different concentrations of BA were studied. Medium containing 1.0 mg l⁻¹ BA and 0.01 mg l⁻¹ IBA was the best, although medium containing 0.6 and 0.8 mg l⁻¹ BA were also successful, and utilisation of 0.4 mg l⁻¹ BA and 0.01 mg l⁻¹ IBA was optimum for shoot elongation. Application of different kinds of auxins (IAA, IBA and NAA at 0.01 or 0.1 mg l⁻¹) with 1.0 mg l⁻¹ BA were also studied. Media containing IBA were significantly better than media containing IAA for shoot fresh weight, but neither of them was significantly different from media containing NAA. Application of 0.01 mg l⁻¹ or 0.1 mg l⁻¹ auxin, with 1.0 mg l⁻¹ BA, was not significantly different for shoot multiplication of Persian walnut. The morphology of shoots on media containing 0.01 mg l⁻¹ IBA was the best.     

Effect of cutting date and position on rooting ability and fatty acid composition of Carignan (Vitis vinifera L.) shoot

The effects of cutting date and position (apical, basal and central) on rooting ability and fatty acid composition from Carignan (Vitis vinifera L.) shoot were determined. Root number and weight depended of the cutting date and position. Only in the case of the cutting in the basal position, there was a highly positive correlation between number and percentage of roots (r = 0.95) during sampling date. Concerning the influence of the cutting date, the root number by cutting oscillated in a saw tooth. The root weight and percentage showed a positive correlation for the three cutting positions and they increased with time. Cutting date and position had an irregularly effect on the contents of total lipid and different fatty acids. Independently of cutting position, the contents of oleic acid (C18:1) and palmitic acid (C16:0) correlated negatively with those of linoleic (C18:2) and linolenic acids (C18:3) during sampling date.     

影响早熟油桃茎尖培养增殖效率的因素

以早熟油桃华光、曙光为试材,研究了外源植物生长调节剂种类及浓度组合、培养基种类、暗培养时间等因素对茎尖培养增殖效率和试管苗生根的影响。结果表明:不同浓度的TDZ和KT对华光、曙光油桃增殖效应不同,在G培养基中加入TDZ2.0mg/L和KT1.5mg/L处理效果最好。1/2MS培养基中附加IBA有利于华光、曙光汕桃的生根;华光油桃生根不必经过暗培养;曙光经过6d或9d暗培养能大大提高生根率和有效根数量。     

Nutritional and cultural factors affecting rooting of papaya (Carica papaya L.) in vitro

Summary

Experiments were conducted to optimize nutritional and cultural requirements for initiation and growth of roots on papaya in vitro. Axillary shoots were obtained from plants which had been sub-cultured monthly for two years. Root initiation was enhanced when 1 to 2 mm of stem base was removed and shoots were growing actively before transfer to the rooting medium. Decreasing daylength during incubation from 24 h to 12 h promoted root initiation. Within the day temperature range of 22 to 29°C, optimum rooting occurred at 27°C and higher temperatures produced higher mean root weights per shoot. High concentrations of growth factors and the absence of sucrose in the medium both reduced root initiation, however, varying the concentration of sucrose and removing growth factors affected mean root weight per shoot. All media contained a modified de Fossard et al. (1974) basal medium plus 10 μM IBA.     

In vitro shoot multiplication and plant regeneration in four Capsicum species using thidiazuron

A procedure of in vitro plant propagation using shoot meristem explants (∼0.5 cm) has been developed for Capsicum annuum cv CA960, C. baccatum, C. frutescens and C. praetermissum on Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Plant Physiol. 15, 473–497] medium containing various cytokinins. Among various concentrations of cytokinins tested; adenine (Ad), N⁶-benzyladenine (BA), kinetin (Kn), zeatin and thidiazuron (TDZ) individually. TDZ regenerated maximum number (4.2–22.4) of shoots in all the Capsicum species tested. Multiple shoot elongation occurred upon transfer to BA (0.22 μM l⁻¹) + IAA (0.48 μM l⁻¹). Rooting of regenerated shoots was achieved on medium supplemented with 5.71 μM l⁻¹ indole-acetic acid (IAA). Rooting was observed in 72–94% of shoots obtained from TDZ-containing regeneration medium followed by elongation treatment in contrast to 8–22% of shoots without elongation treatment. Plantlets obtained from TDZ-containing media were normal diploid (2n = 24) and could readily be established in the soil under green house conditions with a survival frequency of 68–84%. Regenerated plants were developed into morphologically normal, fertile plants and able to set viable seeds.     

Influence of FeEDDHA on in vitro rooting and acclimatisation of red raspberry (Rubus idaeus L.) in peat and vermiculite

Summary

We investigated the dependence of in vitro rooting and acclimatisation to greenhouse conditions on the source of iron used in the shoot multiplication and rooting media using five raspberry (Rubus idaeus L.) cultivars (‘Beskid’, ‘Canby’, ‘Malling Seedling’, ‘Norna’, and ‘Veten’). Ethylenediamine di-2-hydroxy-phenyl acetate ferric (FeEDDHA) in the rooting medium led to higher chlorophyll contents, earlier and more abundant rooting (8.7 vs. 5.3 roots per shoot), 30% higher fresh and dry weights, and thus higher quality microplants than ethylenediaminetetra-acetate ferric sodium (FeEDTA). Higher quality microshoots had a beneficial effect on acclimatisation (i.e., percentage survival and length of shoots) when the microplants were planted in a peat-based substrate; however, when planted in vermiculite, the initial differences disappeared during a 4 week-long growth period in a greenhouse.     

High efficiency shoot regeneration from calluses of strawberry (Fragaria X ananassa Duch.) stipules of in vitro shoot cultures

A highly efficient method of shoot regeneration was developed from calluses of four culti- vars of strawberries (Fragaria x ananassa Duch.), ‘Addie’, ‘Dana’, ‘Gea’ and ‘Santana’, grown in vitro. Optimum shoot regeneration (84-96% of the explants), with four to eight shoots was obtained from calluses developed from stipules near the internal zone between petiole and stipule, on Murashige and Skoog (1962) or Gamborg et al. (1968) media, supplemented with 3% (w/v) glucose, 10 [iM BAP and 2.5 IBA and 0.8% agar. The calluses continued to produce shoots for at least six subsequent subcultures. This has been reported, up to now, only in juvenile explants. Microscopic observation showed no preformed buds or meristematic groups of cells in the connecting zone of petiole and stipule prior to culture. However, there were several layers of cells in this area containing higher amounts of starch, which were not observed in the cells of the bottom or in the external side of the stipule. Regeneration from stipules occurred five to six days earlier in whole leaves (stipule + petiole + lamina) than in leaves without laminae, but the final percentage was the same in the cases of all explants. The percentage of regenerating calli from the other explant sources (leaf lamina, petiole and root) was low and dependent on cultivar. Cv. Gea, which showed the highest regeneration capacity, regenerated 32% from leaf laminae, 16% from petiole and root calluses, followed by cv. Addie with 12% from leaf laminae only; the others failed to regenerate from calluses derived from these tissues. The regenerated shoots were successfully rooted and hardened for further observations on eventual somaclonal variation.     

黑色培养基促进中国樱桃试管苗生根的研究

以中国樱桃抗病毒优系39号试管苗为试材，进行了在培养基中滴厍碳素墨水促根的试验，结果表明，不同蔗糖浓度、不同IBA浓度条件下碳素墨水皆有十分明显的促根作用，最佳组合为1/2MS IBA0.2-0.7mg.L^-1 蔗糖1%，每40-50mg培养基中滴加1滴碳素墨水（约0.04mdl)。     

Medium-term conservation of redwood (Sequoia sempervirens (D. Don.) Endl.) in vitro shoot cultures and encapsulated buds

This study evaluated the survival and recovery of non-encapsulated and encapsulated shoots of Sequoia sempervirens after storage at 4 °C in the dark for up to 15 months on four different culture media. Survival and regrowth of encapsulated shoots declined within 3 months, regardless of the storage medium composition. By contrast, no significant decrease in survival and regrowth was noted with non-encapsulated shoots after 12 months of storage on Quoirin and Lepoivre medium supplemented, or not, with 1 mg l⁻¹ benzyladenine. Regrowth dropped to 60–61% after 15 months of storage on the same media. Medium-term conservation of S. sempervirens germplasm is therefore possible using in vitro storage of non-encapsulated shoot cultures.     

Restoration of rooting competence in a mature plant of Garcinia indica through serial shoot tip grafting in vitro

Apical and axillary buds from a high yielding, early fruiting elite tree (more than 20 years old) were cultured in woody plant medium (WPM) supplemented with 0.9 μM N⁶-benzyl adenine (BA). Multiple shoots were obtained on WPM basal medium containing 8.9 μM BA and 0.5 μM thidiazuron (TDZ). Elongation of axillary shoots was obtained in half-strength WPM medium supplemented with 0.4 μM BA. For root initiation, the elongated shoots were transferred to half strength WPM basal medium containing 2.5–245 μM indole-3-butyric acid (IBA) or 2.7–268.5 μM α-naphthaleneacetic acid (NAA) or the shoots were subjected to 2.5–53.9 mM IBA, 2.7–59.1 mM NAA dip for (30 s–30 min) and then transferred to half strength WPM basal medium. However, rooting was never achieved even after 2 months of culture.     

释放钝绥螨控制柑桔害螨效果试验

顺昌县位于闽北的南部 ,柑桔种植面积 6 6 6 6 6 7ｈｍ2 ,年产量 8万ｔ,为闽北柑桔生产大县 ,柑桔产业是我县农村经济收入的主要来源。红蜘蛛、锈壁虱是当地柑桔重要害虫 ,严重影响柑桔生产的经济效益。为此 ,果农大量使用化学农药进行防治 ,年喷药7～ 11次 ,高者达 13次 ,造成防治成本增加 ,害螨抗药性增强 ,环境受到污染 ,果实农药残留加大 ,天敌大量死亡 ,害螨再增猖獗为害严重。为适应市场经济和加入ＷＴＯ的需要 ,改变长期依赖化学农药造成的不良后果 ,大力推广“以螨治螨”生物防治技术 ,发展优质、高效、无公害生态农业已势在必行。…     

茂谷橘橙引种表现与栽培技术要点

茂谷橘橙原产美国,由佛罗里达州迈阿密农业试验所育成,系宽皮橘与甜橙的杂交种,是佛罗里达州的主栽品种之一。台湾大学园艺系将该品种引入台湾嘉义以北地区试种。福建省农业科学院农业工程技术研究所于1996年从台湾引进茂谷橘橙,高接在漳州长泰8年生桠柑树上,1998年开始结果,     

云南西双版纳大翼厚皮橙的研究

详细报道了生长在云南西双版纳的大翼厚皮橙(Citrus macroptera var.kerrii Swing.)的分布、植物学性状、花粉形态、果实营养成分、果皮精油含量以及种子萌发特点等方面的研究结果。     

In vitro shoot proliferation and enhancement of rooting for the large-scale propagation of yellow bamboo (Bambusa vulgaris ‘Striata’)

Continuous and rapidly proliferating axillary shoots were raised from axillary buds in secondary branches of adult field culms and nursery grown 1-year-old tissue culture-raised plants of Bambusa vulgaris ‘Striata’. Shoots continuously proliferated in a MS medium containing 4 mg L⁻¹ 6-benzyladenine (BA). The effects of indole butyric acid (IBA) levels, a pretreatment with thidiazuron (TDZ) (1-phenyl-1-([1,2,3-thidiazol-5-yl])urea) and illumination on rooting, were investigated after 6 months of shoot proliferation. A rooting medium with IBA at 3 mg L⁻¹ was optimum for root induction. Shoots of adult field culms that were proliferated in the presence of BA when induced to root in this medium resulted in 40% rooting in 27 days. In vitro shoots raised from 1-year-old tissue cultured plants showed 92% rooting under the same conditions. Rooting was enhanced when the relatively difficult-to-root in vitro shoots from adult field culms were pretreated with 0.5 mg L⁻¹ TDZ for two to three subcultures before placing in the root induction medium. Continuously illuminated shoots pretreated with TDZ for three subcultures showed 100% rooting compared to 83% rooting of shoots that were exposed to a 12 h photoperiod. These findings have been applied in the large-scale propagation of this species.     

Root influence on shoot development in tea (Camellia sinensis (L.) O. Kuntze) following shoot pruning

The physiological interaction between the factors controlling the development of new shoots in pruned tea plants was investigated in an experiment using a widely cultivated TRI clone growing at low elevation in Sri Lanka. The functional importance during new shoot growth of root reserves, feeder roots and leaves and buds on unpruned branches (lung-shoots) is critically evaluated. A scheme for their interaction is presented, which highlights the need to maintain feeder roots alive during new shoot development, for which a large proportion of both reserve and current assimilates seem to be utilized. Mature leaves on lung-shoots, besides supplementing carbohydrate supply by current photosynthesis, may also serve to maintain the flow of metabolites from feeder roots to developing shoots and also function as ‘sinks’ for excess, toxic levels of root metabolites. Removing all mature leaves, as is done during ‘clean pruning’, leads to the depletion of reserves below the critical level for feeder root survival, resulting in death of feeder roots and failure of new buds to form shoots.     

紫外线照射对梁平柚果皮基因表达的影响

 采用SSH技术以紫外照射的梁平柚[Citrus grandis（L.）Osbeck]果皮作为实验方（tester）,以未被照射的正常果皮作为驱动方（driver）,构建了一个梁平柚果皮紫外诱导基因的正向差减文库。经菌液PCR检测后随机挑取200个阳性克隆测序,获得168条表达序列标签（ESTs）。比对这168条ESTs,发现有分属于57个基因的114条ESTs与已知基因高度同源,54条ESTs同源性较低或没有同源性。功能分析发现,这些ESTs主要参与抗逆防御、生长发育、细胞凋亡、转录与翻译、细胞分化、信号传导、能量代谢、糖类及氨基酸代谢以及次生代谢等。     

Effect of gibberellic acid (GA3) on elongation and rooting of ‘St. Julien A’ rootstock in vitro

‘St. Julien A’ (Prunus instititia L.) rootstock was induced to proliferate shoots on a modified half-strength Murashige and Skoog (MS) medium. Cultures treated with 12.5 mg l^?1 gibberellic acid (GA₃) produced elongated shoots suitable for rooting. Elongated shoots were placed in media with indolebutyric acid (IBA) or indole-3-acetic acid (IAA) with or without a 16-day dark incubation. Light (16-h photoperiod) inhibited rooting. IAA (4 mg l^?1) was ineffective in promoting rooting. Rooting was best when shoots were incubated in the dark with IBA (4 mg l^?1). GA₃ was deleterious to shoots, causing chlorosis and apical die-back. Light regime interacted with auxin treatments in affecting shoot condition. Shoot condition was better on shoots treated with IBA and dark-incubated; while those treated with IAA were better when light-incubated.     

采后钙及热处理对葡萄柚果实贮藏期细胞壁物质代谢的影响

以“里约红”葡萄柚果实为试材,用3％ CaCl2、50℃热水单独、复合浸泡处理5 min,在 室温(18±2)℃、相对湿度85％～90％的环境条件下贮藏,研究采后钙、热处理对葡萄柚果实的贮藏效果及细胞壁物质代谢的抑制作用.结果表明:采后钙、热处理有效控制了果实硬度下降、失重率上升,维持果实共价结合型果胶(SCSP)、24％半纤维素(24KSF)含量,降低了果实水溶性果胶(WSP)、果实离子型果胶(ESP)、4％果实半纤维素(4KSF)含量.其中,以3％CaCl2+50℃处理效果最佳,贮藏75 d果实硬度、乙醇不溶物、果实共价结合型果胶(SCSP)、24KSF分别比对照高出21.31％、6.44％、22.60％和50.46％.此外,钙处理效果优于50℃热处理.相关性分析结果表明,果实硬度与细胞壁各组分呈显著相关.通过主成分分析构建的综合评价模型可知,采后钙、热处理可提高贮藏期间葡萄柚果实质地综合品质,抑制细胞壁物质代谢降解,从而延长葡萄柚果实贮藏期.