三种生物源农药对平菇双翅目害虫的药效试验

研究了苦皮藤素、印楝素和苏云金杆菌以色列变种(Bti)3种生物源农药对平菇双翅目害虫的防治效果。大棚药效试验表明,0.6%印楝素乳油的防治效果最好,其次为1%苦皮藤素乳油和Bti,2次施药5 d后三种药剂防效分别为58.7%、47.5%和26.9%,2次施药后10 d的防效分别为74.3%、58.6%和42.5%。三种生物源药剂对平菇子实体的形成和产量不产生影响。     

10％世高防治梨黑斑病药效试验

用10％世高6000倍液、50％多菌灵800倍液、70％甲基托布津1000倍液、50％先锋1500倍液等药剂进行梨黑斑病的田间防治试验。试验结果表明：10％世高6000倍液和70％甲基托布津1000倍液对梨黑斑病防治效果显著高于其他药剂，全年喷药5次，其防治效果分别达到82．34％和78．63％。为了降低防治成本，生产上可用10％世高和70％甲基托布津交替使用防治梨黑斑病。     

4种杀虫剂对苹果蚜虫的田间防治效果

为筛选防治苹果蚜虫安全有效的杀虫剂,采用田间试验的方法,研究了21%噻虫嗪悬浮剂5000倍液、5%啶虫脒乳油2500倍液、5%吡虫啉可溶液剂2000倍液和2.5%溴氰菊酯乳油2000倍液4种杀虫剂对苹果蚜虫的田间防治药效。试验结果表明:4种药剂处理对苹果树蚜虫的防效均达70%以上,具有良好的防治效果,在试验用量下苹果树未出现药害和其它不良反应。     

防治豇豆蚜虫和美洲斑潜蝇的田间药效试验

选用2.5%吡虫啉乳油、2.5%高效氯氟氰菊酯乳油、5%啶虫脒乳油、100g.L-1高效灭百可乳油、25%噻嗪酮可湿性粉剂对豇豆蚜虫进行田间防治试验;选用480g.L-1毒死蜱乳油、4.5%高效氯氰菊酯乳油、80%敌敌畏乳油、75%灭蝇胺可湿性粉剂和0.5%甲氨基阿维菌素苯甲酸盐乳油对豇豆美洲斑潜蝇进行田间防治试验。结果表明:药后1～10d,2.5%吡虫啉乳油1500倍液、2.5%高效氯氟氰菊酯乳油2000倍液、5%啶虫脒乳油1000倍液和100g.L-1高效灭百可乳油2000倍液对豇豆蚜虫防治效果均在95.76%以上,防效优良;25%噻嗪酮可湿性粉剂1000倍液对豇豆蚜虫的防治效果仅为29.79%～66.68%,防效很差。75%灭蝇胺可湿性粉剂3000倍液对美洲斑潜蝇防效最好,药后3、7和10d的防治效果分别为89.63%、94.10%和93.28%;其次是480g.L-1毒死蜱乳油1000倍液,药后3、7和10d的防治效果分别为61.12%、69.15%和69.40%;其他几种药剂的防效均较差。     

1%苦皮藤素水乳剂防治黄曲条跳甲田间试验

推广应用高效、低毒、低残留的生物农药是保障蔬菜产品质量安全的重要途径。为验证1%苦皮藤素水乳剂对青菜黄曲条跳甲的防治效果,作者开展了相关试验。试验结果表明,1%苦皮藤素水溶剂各浓度处理均可减轻黄曲条跳甲成虫的危害程度,在试验剂量下对青菜生长未产生不良影响,对周边环境友好;从防效和防治持效期来看,1%苦皮藤素水乳剂500倍液的防效较理想,5%鱼藤酮可溶液剂300倍液发挥药效较慢,但持续期较长。     

樱桃园苹毛丽金龟发生规律及防治

苹毛丽金龟 (Proagopertha lucidula Fal.)又叫苹毛金龟子 ,俗称铜克郎 ,属鞘翅目金龟总科 ,是北方果树花期主要害虫 ,危害多种果树及经济林木。顺义樱桃园为山地果园 ,环境复杂 ,园内果树品种甚多 ,多年来苹毛丽金龟一直是主要害虫 ,每年损失严重 ,尤其是经济效益较高的樱桃园 ,几乎连年绝产 ,却一直没有有效的防治措施 ,为此 ,2 0 0 0年对苹毛丽金龟的越冬场所、活动规律和食性进行了研究 ,并以此为基础 ,采取了一系列防治措施 ,为无公害果品的生产奠定基础。1　材料与方法1 .1　果园条件果园地处北京东北郊顺义区赵各庄乡 ,果园西北东三…     

防治桃蚜虫的药剂筛选试验

通过对桃蚜虫药剂筛选试验,结果表明,供试的4种药剂,对桃树蚜虫均有良好的防治效果。以5%马拉松乳剂1000倍夜的防治效果最佳,施药后2d、5d、8d的防效分别为72.03%、93.73%、95.74%。与其余3种药剂的防效相比,有显著性差异。     

五种生物农药对阿克苏地区苹果叶螨室内毒力及田间药效试验

以山楂叶螨(Tetranychus viennensis)为试材,采用玻片浸渍法测定4种生物农药对山楂叶螨的室内毒力,并在田间评价了5种生物农药对山楂叶螨的防治效果。结果表明:室内毒力测定试验中1%苦皮藤素对雌成螨LC50为0.826 3 mg·L~(-1);其次为1%苦参·印楝素和1.5%除虫菊素,LC50分别为0.999 5mg·L~(-1)和5.756 0mg·L~(-1);40%阿维菌素·矿物油毒性最低,LC50为8.765 7mg·L~(-1)。田间防效试验中1%苦参·印楝素1 000倍液、1%苦皮藤素500、1 000倍液、40%阿维菌素·矿物油2 000倍液7d后相对防效都达到88.1%以上。因此,1%苦参·印楝素1 000倍液、1%苦皮藤素500、1 000倍液、40%阿维菌素·矿物油2 000倍液可作为阿克苏地区山楂叶螨防治的首选生物农药。     

梨小食心虫的发生及药剂防治试验

用梨小食心虫性诱芯诱捕成虫,观察梨小食心虫的发生动态并进行喷药防治,结果表明,梨小食心虫在泰安1年发生4～5代,当连续诱得成虫2～3天后喷药防治,35％氯虫苯甲酰胺水分散粒剂10000倍液优于48％乐斯本乳油1500倍液的防治效果,药剂持效期20天以上.     

柑桔粉虱防治药剂的筛选

对13种药剂进行了防治柑桔粉虱的田间药效试验。结果表明,筛选出的防治效果较好、对柑桔安全的药剂有95%机油乳油50倍液、24%BYI8330悬浮剂1000倍液、40%杀扑磷乳油500倍液、40%毒死蜱乳油500倍液、25%扑虱灵可湿性粉剂500倍液和5%啶虫脒乳油1000倍液。供试药剂对柑桔粉虱1龄若虫效果最好,对卵无效。最好在1龄若虫孵化高峰期进行药剂防治。     

Plant growth,yield, and fruit quality of ‘Fallglo’ and ‘Sunburst’ mandarins on four rootstocks

Vegetative growth, yield, and fruit quality of ‘Fallglo’ and ‘Sunburst’ mandarins on ‘Rangpur’ lime, ‘Swingle’ citrumelo, ‘Orlando’ tangelo, and ‘Cleopatra’ mandarin were evaluated under subtropical climate of Northern São Paulo State, Brazil, from 2000 through 2006 harvest seasons. ‘Fallglo’ mandarin trees had the highest cumulative yield on ‘Rangpur’ lime, and the smallest on ‘Swingle’ citrumelo and ‘Orlando’ tangelo. Plants of this cultivar had the highest yield efficiency on ‘Rangpur’ lime, and the lowest on ‘Orlando’ tangelo. ‘Sunburst’ mandarin trees began to bear fruits later than ‘Fallglo’ mandarin trees, with no differences in yield induced by the rootstocks. ‘Cleopatra’ mandarin induced the most vigorous growth in ‘Fallglo’ mandarin as compared to plants on ‘Swingle’ citrumelo. On the other hand, the largest trees of ‘Sunburst’ mandarin were registered on ‘Orlando’ tangelo, and the smallest on ‘Rangpur’ lime. ‘Sunburst’ mandarin had higher alternate bearing than ‘Fallglo’ regardless the rootstock. Fruit weight and juice content were not affected by the rootstock. These two mandarin scion cultivars may be considered adequate alternatives to produce good fruit quality for the fresh fruit market. ‘Cleopatra’ mandarin and ‘Rangpur lime are suitable rootstocks for ‘Fallglo’ mandarin, whereas all rootstocks evaluated are adequate for ‘Sunburst’ mandarin.     

Tree performance and fruit yield and quality of ‘Okitsu’ Satsuma mandarin grafted on 12 rootstocks

The citriculture in Brazil, as well as in other important regions in the world, is based on very few mandarin cultivars. This fact leads to a short harvest period and higher prices for off-season fruit. The ‘Okitsu’ Satsuma (Citrus unshiu Marc.) is among the earliest ripening mandarin cultivars, and it is considered to be tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.) and to citrus variegated chlorosis (Xylella fastidiosa Wells et al.). Despite having regular fruit quality under hot climate conditions, the early fruit maturation and absence of seeds of ‘Okitsu’ fruits are well suited for the local market in the summer (December through March), when the availability of citrus fruits for fresh consumption is limited. Yet, only a few studies have been conducted in Brazil on rootstocks for ‘Okitsu’. Consequently, a field trial was carried out in Bebedouro, São Paulo State, to evaluate the horticultural performance of ‘Okitsu’ Satsuma mandarin budded onto 12 rootstocks: the citrandarin ‘Changsha’ mandarin (Citrus reticulata Blanco) × Poncirus trifoliata ‘English Small’; the hybrid Rangpur lime (Citrus limonia Osbeck) × ‘Swingle’ citrumelo (P. trifoliata (L.) Raf. × Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.) ‘Rubidoux’, ‘FCAV’ and ‘Flying Dragon’ (P. trifoliata var. monstrosa); the mandarins ‘Sun Chu Sha Kat’ (C. reticulata Blanco) and ‘Sunki’ (Citrus sunki (Hayata) Hort. ex. Tanaka); the Rangpur limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (Citrus sinensis × P. trifoliata), ‘Swingle’ citrumelo (P. trifoliata × C. paradisi), and ‘Orlando’ tangelo (C. paradisi × Citrus tangerina cv. ‘Dancy’). The experimental grove was planted in 2001, using a 6 m × 3 m spacing, in a randomized block design. No supplementary irrigation was applied. Fruit yield, canopy volume, and fruit quality were assessed for each rootstock. A cluster multivariate analysis identified three different rootstock pairs with similar effects on plant growth, yield and fruit quality of ‘Okitsu’ mandarin. The ‘Flying Dragon’ trifoliate had a unique effect over the ‘Okitsu’ trees performance, inducing lower canopy volume and higher yield efficiency and fruit quality, and might be suitable for high-density plantings. The ‘Cravo Limeira’ and ‘Cravo FCAV’ Rangpur limes induced early-ripening of fruits, with low fruit quality. ‘Sun Chu Sha Kat’ and ‘Sunki’ mandarins and the ‘Orlando’ tangelo conferred lower yield efficiency and less content of soluble solids for the latter rootstock.     

Identification of S-alleles in pear (Pyrus communis L.) cv. ‘Rocha’ and other European cultivars

In this work we report the cloning and identification of S-RNase alleles responsible for gametophytic self-incompatibility (GSI) of ‘Rocha’ pear and of 13 other European pear cultivars that might be used as its pollinators. Partial sequences of S-RNase alleles were amplified by PCR with specific primers hybridising in conserved regions of previously identified S-RNase alleles of Pyrus communis, cloned and sequenced and the S-genotype of eight pear cultivars was fully determined. Three cultivars (‘General Léclerc’ (SqSl), ‘Tosca’ (SbSl) and ‘Alexandrine Douillard’ (SbSk)) shared no S-alleles with ‘Rocha’ (SaSe) and shall be totally compatible with this cultivar. None of the cultivars analysed showed an identical amplification pattern to the one observed in ‘Rocha’, so the other cultivars shall be at least semi-compatible. One new allele was identified in P. communis cv. ‘Beurré d’Avril’ (designated as St). The determination of both S-RNase alleles of cvs ‘Rocha’, ‘Beurré Precoce Morettini’ (SeSk) and ‘Tosca’ and the identification of one S-RNase allele in cvs ‘Carapinheira’ (Sb), ‘Amêndoa’ (Se), ‘Pérola’ (Sk) and ‘Beurré d’Avril’ (St) are important contributions for the effort recently developed worldwide to establish groups of sexual compatibility among European pears.     

Evaluation of citrus somatic hybrids for tolerance to Phytophthora nicotianae and citrus tristeza virus

Somatic hybridization is a biotechnology tool that can be used in citrus breeding programs to produce somatic hybrids with the complete genetic combination of both parents. The goal of this work was to test the reaction of citrus somatic hybrids that may be useful as rootstocks to trunk and root infections caused by Phytophthora nicotianae van Breda de Haan (P. parasitica Dastur) and to citrus tristeza virus (CTV). The somatic hybrids evaluated were ‘Caipira’ sweet orange (Citrus sinensis L. Osbeck) + ‘Rangpur’ lime (C. limonia Osbeck), ‘Caipira’ sweet orange + ‘Cleopatra’ mandarin (C. reshni hort. ex Tanaka), ‘Caipira’ sweet orange + ‘Volkamer’ lemon (C. volkameriana V. Ten. & Pasq.), ‘Caipira’ sweet orange + rough lemon (C. jambhiri Lush.), ‘Cleopatra’ mandarin + ‘Volkamer’ lemon, ‘Cleopatra’ mandarin + sour orange (C. aurantium L.), ‘Rangpur’ lime + ‘Sunki’ mandarin (C. sunki (Hayata) hort. ex Tanaka), ‘Ruby Blood’ sweet orange (C. sinensis L. Osbeck) + ‘Volkamer’ lemon, ‘Rohde Red’ sweet orange (C. sinensis L. Osbeck) + ‘Volkamer’ lemon, and ‘Valencia’ sweet orange + Fortunella obovata hort. ex Tanaka. For P. nicotianae trunk and root infection assays, plants of the somatic hybrids, obtained from 9-month semi-hardwood cuttings, were evaluated and compared with diploid citrus rootstock cultivars after mycelia inoculation in the trunk or spore infestation in the substrate, respectively. ‘Cleopatra’ mandarin + sour orange, ‘Rangpur’ lime + ‘Sunki’ mandarin, ‘Cleopatra’ mandarin + ‘Volkamer’ lemon, ‘Ruby Blood’ sweet orange + ‘Volkamer’ lemon, ‘Rohde Red’ sweet orange + ‘Volkamer’ lemon, and ‘Caipira’ sweet orange + ‘Volkamer’ lemon had less trunk rot occurrence, whereas the somatic hybrids ‘Cleopatra’ mandarin + ‘Volkamer’ lemon, ‘Cleopatra’ mandarin + sour orange, ‘Caipira’ sweet orange + ‘Volkamer’ lemon, and ‘Caipira’ sweet orange + ‘Rangpur’ lime were tolerant to root rot. For CTV assays, plants of the somatic hybrids along with tolerant and intolerant rootstocks were budded with a mild strain CTV-infected or healthy ‘Valencia’ sweet orange budwood. Differences in average scion shoot length indicated that the hybrids ‘Cleopatra’ mandarin + sour orange and ‘Valencia’ sweet orange + Fortunella obovata were intolerant to CTV.     

Horticultural performance of ‘Folha Murcha’ sweet orange onto twelve rootstocks

Despite its outstanding position, the Brazilian citriculture is established on a very limited pool of varieties that limits its expansion and restricts the fruit availability throughout the year. This situation determines the urgent necessity of developing alternative scion and rootstock cultivars, with good performance under local conditions. ‘Folha Murcha’ sweet orange (Citrus sinensis (L.) Osbeck) is a late-harvest cultivar, suitable both for the juice processing industry and the fresh fruit market, being described as tolerant to citrus canker (Xanthomonas citri subsp. citri Schaad et al.), and less affected by citrus variegated chlorosis (Xylella fastidiosa Wells et al.). A study was conducted in Bebedouro, São Paulo State, Brazil, to evaluate the horticultural performance of ‘Folha Murcha’ sweet orange budded onto 12 rootstocks: the citrandarin ‘Changsha’ mandarin (Citrus reticulata Blanco) × Poncirus trifoliata ‘English Small’; the hybrid ‘Rangpur’ lime (Citrus limonia Osbeck) × ‘Swingle’ citrumelo (P. trifoliata (L.) Raf. × Citrus paradisi Macfad.); the trifoliates (P. trifoliata (L.) Raf.) ‘Rubidoux’, ‘FCAV’, and ‘Flying Dragon’ (P. trifoliata var. monstrosa); the ‘Sun Chu Sha Kat’ mandarin (C. reticulata Blanco); the ‘Sunki’ mandarin (Citrus sunki (Hayata) Hort. ex. Tanaka); the ‘Rangpur’ limes (C. limonia Osbeck) ‘Cravo Limeira’ and ‘Cravo FCAV’; ‘Carrizo’ citrange (C. sinensis × P. trifoliata), ‘Swingle’ citrumelo (P. trifoliata × C. paradisi), and ‘Orlando’ tangelo (C. paradisi × Citrus tangerina cv. ‘Dancy’). The experimental grove was planted in 2001, using a 7 m × 4 m spacing, in a randomized block design, with five replications and two plants per plot. No supplementary irrigation was applied. Fruit yield, canopy volume, tree tolerance to drought and to citrus variegated chlorosis, and fruit quality were assessed for each rootstock. Trees grafted onto the ‘Flying Dragon’ trifoliate were smaller in size, but had largest yield efficiency when compared to those grafted onto other rootstocks. Lower alternate bearing index was observed on trees budded onto ‘Cravo FCAV’ ‘Rangpur’ lime. Both ‘Rangpur’ lime rootstocks and the ‘Sunki’ mandarin induced higher tree tolerance to drought. The ‘Flying Dragon’ trifoliate induced better fruit quality and higher tolerance to citrus variegated chlorosis (CVC) to ‘Folha Murcha’ trees. A cluster multivariate analysis identified three groups of rootstocks with similar effects on ‘Folha Murcha’ tree performance. Among the 12 evaluated rootstocks, the ‘Flying Dragon’ trifoliate has a unique effect on plant growth, tolerance to drought and CVC, fruit yield and fruit quality of ‘Folha Murcha’ trees, and may be better suited for high-density plantings.     

Effect of 2,4-d, glutamine and BAP on embryogenic suspension culture of date palm (Phoenix dactylifera L.)

The proliferation of embryogenic suspension culture in two cultivars (Jihel and Bousthami Noir) of Phoenix dactylifera L. was tested on liquid media with or without 2,4-d and with different glutamine concentrations (3.35 × 10⁻⁴, 6.7 × 10⁻⁴ and 13.4 × 10⁻⁴ M). The liquid medium with 0.1 mg l⁻¹ 2,4-d and 6.7 × 10⁻⁴ M glutamine has clearly improved the proliferation of somatic embryos. In fact, when glutamine concentration increased from 3.35 × 10⁻⁴ to 6.7 × 10⁻⁴ M, the yield of somatic embryos increased from 14 to 56 embryos per 100 ml of culture medium for “Jihel” cultivar and 25–71 embryos per 100 ml of culture medium for “Bousthami Noir” cultivar. In contrast, increasing glutamine concentration from 6.7 × 10⁻⁴ to 13.4 × 10⁻⁴ M, the embryos yield was negligible. Based on biochemical analysis, the highest accumulation of proteins and sugars was obtained in liquid medium with 0.1 mg l⁻¹ 2,4-d and 6.7 × 10⁻⁴ M glutamine (118 and 91 mg of proteins g⁻¹ DW, respectively, for “Jihel” and “Bousthami Noir” cultivars; 194 mg of sugars g⁻¹ DW for “Jihel” cultivar and 182 mg of sugars g⁻¹ DW for “Bousthami Noir” cultivar). In addition, the supply of 0.05 mg l⁻¹ BAP on the germination medium could be useful in terms of germination percentage of somatic embryos. When BAP concentration increased from 0.05 to 0.2 mg l⁻¹, the germination percentage of somatic embryos decreased from 14.2 to 4.9%, while secondary embryogenesis increased from 26.4 to 45.2%.     

Changes in fox squirrel anti-predator behaviors across the urban–rural gradient

Predator stimuli created by humans in the urban environment may alter animals’ anti-predator behaviors. I hypothesized that habituation would cause anti-predator behaviors to decrease in urban settings in response to humans. Additionally, I hypothesized that populations habituated to humans would show reduced responses to other predator stimuli. I observed three populations of squirrels (urban, suburban and rural) responses to human approaches, red-tailed hawk vocalizations (Buteo jamaicensis) and coyote (Canis latrans) vocalizations. Mahalanobis distances of anti-predator behaviors in response to human approaches were consistent with the urban–rural gradient. Flight initiation distances (X ² = 26.33, df = 2, P < 0.001) and amount of time dedicated to anti-predator behavior (X ² = 10.94, df = 2, P = 0.004) in response to human approaches were also consistent with the urban–rural gradient. Supporting the habituation hypothesis, naive juvenile squirrels increased flight initiation distances (X ² = 35.89, df = 1, P < 0.001) and time dedicated to anti-predator behaviors (X ² = 9.46, df = 1, P = 0.002) relative to adult squirrels in the same urban environment. Time dedicated to anti-predator behaviors differed among all three sites in response to both coyote (X ² = 9.83, df = 2, P = 0.007) and hawk (X ² = 6.50, df = 2, P = 0.035) vocalizations. Responses to both vocalizations on rural sites (coyote = 45%, hawk = 55%) greater than twice that found on the urban sites (coyote = 11%, hawk = 20%). This is possibly the first case of a transfer of habituation demonstrated under field conditions.     

Production of inter-section hybrids between Primula filchnerae and P. sinensis through ovule culture

Inter-section hybrids were obtained in the reciprocal crosses between Primula filchnerae (2n = 2x = 24) of Sect. Pinnatae and P. sinensis ‘Fanfare’ (2n = 2x = 24) of Sect. Auganthus by rescuing ovules on half-strength (1/2) Murashige and Skoog's (MS) medium supplemented with 50 g l⁻¹ sucrose, 2.5 g l⁻¹ gellan gum, 0.1 mg l⁻¹ α-naphthaleneacetic acid (NAA), 0.1 mg l⁻¹ 6-benzyladenine (BA) and 50 mg l⁻¹ gibberellic acid (GA₃). In ovule culture, germination occurred with radicle elongation but no plumule was observed. The radicle kept on the initial medium showed root proliferation with callus formation. When the calluses were transferred to (1/2)MS media containing 30 g l⁻¹ sucrose and 3 g l⁻¹ gellan gum, without plant growth regulators (PGRs) or with 1 mg l⁻¹ zeatin and 0.1 mg l⁻¹ NAA, plantlets were regenerated. The plants thus obtained were confirmed to be hybrids through flow cytometry (FCM) and random amplified polymorphic DNA (RAPD) analyses. The hybrid obtained when P. filchnerae was used as the maternal parent was diploid, whereas hexaploid hybrid was obtained when using P. sinensis as the maternal parent. The hexaploid hybrid might be produced through chromosome doubling of a triploid originated from the fertilization of P. sinensis with unreduced pollen of P. filchnerae.     

Plant regeneration of pansy (Viola wittrockiana) ‘Caidie’ via petiole-derived callus

An in vitro plant regeneration protocol for pansy (Viola wittrockiana) cultivar ‘Caidie’ from petioles was established as following: callus induction on a half-strength MS medium supplemented with 0.45 μmol l⁻¹ 2,4-d plus 8.9 μmol l⁻¹ BA, callus subculture on medium F (1/2MS with 4.5 μmol l⁻¹ 2,4-d, 2.7 μmol l⁻¹ NAA and 0.44 μmol l⁻¹ BA) and then on medium T (1/2MS with 4.5 μmol l⁻¹ 2,4-d, 2.7 μmol l⁻¹ NAA and 2.2 μmol l⁻¹ BA), shoot regeneration on medium D3 (MS media supplemented with 2.9 μmol l⁻¹GA₃, 23.6 μmol l⁻¹ AgNO₃, 0.02% active charcoal and 4.5 μmol l⁻¹ TDZ), shoot multiplication on medium M (half-strength MS medium containing NAA 1.1 μmol l⁻¹, TDZ 9.1 μmol l⁻¹ and GA₃ 8.7 μmol l⁻¹), and then shoot elongation and rooting on medium R (MS medium supplemented with 1.1 μmol l⁻¹ NAA and 1.1 μmol l⁻¹ BA). Subculture on appropriate medium was found to be important for successful shoot regeneration.