Preliminary results of using in vitro axillary and adventitious buds in mutation breeding of Chinese gooseberry

Summary Aseptically cultured shoots of Chinese gooseberry exhibited growth disorder and morphological aberrances, and some died after being exposed to sufficient gamma-ray irradiation. The death rate was dose dependant and the LD₅₀ was 80–90 Gy and 50–60 Gy respectively for cv. Hayward and clone 4. All petiole explants irradiated with gamma-ray could form calli as the control, but the rate of differentiation of adventitious shoots of the petiole explants decreased and was dependant on dose. Sensitivity of the shoot or petiole explants to gamma-ray irradiation varied with species. Gamma-ray irradiation did not deter either the 2-node segments from producing axillary shoots M₁, M₂, and M₃ or the advantitious shoots originating in the petiole explants and the M₃ shoots from forming advantitious roots. Therefore, using aseptically cultured axillary or adventitious buds for mutation breeding of Chinese gooseberry is feasible. A bacterium surviving in the explants lessened the efficiency of these two in vitro techniques in mutation breeding of Chinese gooseberry.Abbreviations IAA 3-indole acetic acid - IBA -indole butyric acid - MS Murashige & Skoog (1962)     

Genetic variation in dry weight of carnation cuttings: its causes and consequences

Summary Carnation cultivars vary considerably in the average weight of their cuttings. Cutting weight shows a high negative correlation with mean relative growth rate. This was initially attributed to differences in age of the cuttings when taken from the stock plants, age being defined as time elapsed since start of visible shoot growth. Shoots on a stock plant are removed as cuttings when they have reached an adequate size and number of leaf pairs. Cuttings from cultivars with a lower rate of growth are removed later, usually resulting in a higher dry weight. This could explain why a high cutting weight is associated with a low relative growth rate.The validity of this tentative explanation is tested in 13 carnation cultivars grown under controlled conditions in two trials, one with plants grown from pinched cuttings and one with plants grown from unpinched cuttings for early flowering (forcing). An analysis of the data for plant weight and leaf pair number at successive stages of development suggests that the primary cause of the negative correlation between cutting weight and mean relative growth rate is not genetic variation in age of the cuttings, but variation in the size of the fully developed axillary bud on the stock plant. Larger buds produce thicker shoots that grow more slowly. Their later harvesting as cuttings only enhances their initial weight advantage, and also the negative correlation with mean relative growth rate.When plants are pinched 20 days after planting, as is normal in a summer-grown crop, the newly developing shoots still show variation in age. This is attributed to variation in the developmental stage of the dormant axillary buds at the time of pinching, causing variation in the time interval between pinching and the start of visible shoot growth.     

茶树不同组织体细胞胚、不定芽分化的研究

根据茶树子叶柄、叶柄断面离腋芽的距离,将外植体依次分为C1, C2, C3和L1, L2, L3。通过上述外植体和不同培养基的研究结果表明:子叶柄外植体在MT1和MT3培养基上培养,其不定芽及总分化率是C1>C2>C3;体细胞胚的分化率是C3>C2>C1。叶柄外植体只有L1在MT3培养基上培养分化了芽,MT1培养基的胚性愈伤组织诱导率比MT,培养基高,     

Vegetative propagation of Beta vulgaris by leaf cuttings with axillary buds

Summary In leaf cuttings with a short piece of hypocotyl tissue axillary bud development was poor. Most cuttings with a long piece of hypocotyl tissue developed into plants. The latter type enabled vegetative propagation of various genotypes.     

驱蚊草组织培养和快速繁殖技术的研究

采用植物组织培养技术,利用驱蚊草顶芽及带腋芽茎段为外植体,开展驱蚊草的诱导分化、芽苗增殖、壮苗培养、生根和移栽等系列研究。结果表明,采用二次消毒方法,有利于无菌系的建立;在无菌系建立后,以带叶柄叶片为材料,在MS+BA0.5mg/L+NAA0.1mg/L培养基上进行培养,可分化、获得高达60%的正常丛生芽;以1/2MS+6-BA0.1mg/L+NAA0.1mg/L为丛生芽增殖继代培养基,不仅能够满足快繁、增殖,同时也能够降低驱蚊草试管苗玻璃比发生频率;生根培养基用1/2MS+6-BA0.1mg/L+NAA0.5mg/L外加AC0.5mg/L,炼苗后将瓶苗移栽到泥炭土、沙、蛭石(质量比为2:3:1)的基质中,控温、保温、保湿栽,成活率可达80%左右。     

彰武松腋芽诱导研究

为了探讨彰武松组培快繁的可行性,以彰武松封顶芽为外植体进行培养,研究灭菌方法、培养基成分以及植物生长调节物质等对其腋芽诱导的影响。结果表明：外植体用70%的乙醇处理30 s后用0.1%的HgCl2灭菌5 min,在添加BA 2~5 mg/L和蔗糖30 g/L的WPM培养基上,腋芽诱导率达50%~70%,NAA对腋芽诱导无促进作用。通过在添加和不添加植物生长调节物质(PGR)的WPM培养基中交替继代培养可以实现芽丛伸长与增殖。     

In vitro production of male sterile Zinnia elegans

Summary Axillary bud proliferation of Zinnia elegans Jacq. was accomplished from vegetative bud explants of greenhouse-grown plants. Photoperiod extension via metalarc lamps in the greenhouse stock plant environment significantly increased both the quantity and quality of explants. Shoot cultures initiated from axillary buds of identified male sterile plants proliferated best on a medium with 1M BAP. Microshoots exhibited 80% rooting on media without growth regulators, and 100% acclimatization of rooted plantlets was accomplished in a lighted, high humidity chamber. In vitro- produced male sterile plants were more compact, more heavily branched, and demonstrated earlier flowering than seed-propagated plants. These male steriles were cross pollinated to male fertile lines to verify their ability to serve as a uniform female parent.     

巴西橡胶树自根幼态无性系的试管微繁

带有腋芽的橡胶花药苗切段接种在MS 6—BA2mg/L的培养基上,30～40天新芽可长至3～5cm高,并可切割再次增殖;切取3～5cm高带叶片的增殖芽作为插条,基部经50～100mg/L IBA预处理再插到生根培养基上,生根率达95％;诱发的根系与茎杆之间愈合良好,输导组织连接紧密,生根植株移栽成活率达85％以上。     

粗肋草‘中国红’植株再生体系的建立

为建立相对稳定、高效的‘中国红’粗肋草植株再生体系,本研究以‘中国红’粗肋草的嫩茎腋芽原基、嫩叶为外植体,对诱导愈伤组织的初代培养基、芽增殖分化培养基的激素种类和配比以及生根培养方法等进行筛选优化。结果表明:适宜建立‘中国红’粗肋草体细胞胚发生途径再生体系的外植体为腋芽原基;最佳诱导愈伤组织的初代培养基为1/2MS+2,4-D 0.2 mg/L+6-BA 0.5 mg/L,愈伤组织诱导率分别为:腋芽原基83.3%,嫩叶为23.9%;适合腋芽原基愈伤组织分化培养基的激素配比为6-BA/IAA组合,愈伤组织分化率为65.7%~67.6%、主要以体细胞胚性途径形成再成芽、芽增殖系数5.3~5.8;适宜的生根接种、培养方式为体细胞胚植株断根促根培养,生根苗生根率99.2%。本研究已初步建立和优化以腋芽原基为外植体的稳定‘中国红’粗肋草体细胞胚发生途径的植株再生体系,可进一步改良优化,为今后‘中国红’粗肋草工厂化及规模化生产提供理论和技术指导。     

枸树组织培养的研究

对枸树的初代培养、继代培养和生根培养进行了研究。初代培养用腋芽、顶芽作为外植体,在灭菌前用半胱氨酸处理10min,以防止褐变。培养出以芽丛为主,以茎段为辅的无性繁殖系。其培养基为MS+BA1.0mg/L+NAA0.1mg/L。继代培养时采用芽丛和茎段进行增殖,其培养基为MS+BA0.5mg/L。在培养基中增加蔗糖的用量和降低光照,可以减轻玻璃化现象。继代苗可以生根,其培养基为1/2MS+IBA0.5 mg/L,但生根的速度较慢。     

几种陆地棉腋芽快速繁殖的研究

以陆地棉的三个品种 (系 )棕色棉 ( X0 0 7)、绿色棉 ( X0 0 8)和白色棉 coker31 2为材料 ,进行不同培养条件下棉花幼苗腋芽快速繁殖的比较试验。分析了不同棉花品种 (系 )基因型、不同培养基类型、以及不同无菌苗苗龄对腋芽快速繁殖的影响因子 ,初步建立了陆地棉腋芽的诱导、伸长、生根、壮苗和移栽的技术体系 ,并将该技术体系在转acs B基因彩色棉品系的扩繁上得到了较理想的应用     

濒危药用植物天山雪莲高效植株再生体系的建立

以濒危药用植物天山雪莲叶片为研究材料进行组织培养和植株再生研究,旨在建立天山雪莲的高效植株再生体系,为其种群复壮、资源保护与可持续利用提供技术参考。研究结果表明:适合天山雪莲叶片愈伤组织诱导的最佳培养基为MS+2,4-D(0.5 mg/L)+6-BA(1.5 mg/L),诱导出愈率为97%;适合愈伤组织丛生芽分化的最佳培养基为MS+NAA(0.05 mg/L)+6-BA(0.4 mg/L)+水解乳蛋白(500 mg/L),分化率为66%,增殖倍数为5.1;适合生根的最佳培养基为1/2 MS+NAA(0.3 mg/L),在此条件下,根发育良好,生根率为76.9%,植株健壮;组培苗炼苗后移栽,成活率可达89%。     

利用横向薄层培养技术建立番木瓜植株再生体系

建立一个高效稳定的植株再生系统是番木瓜生物技术育种和品种改良的关键。本研究通过横向薄层培养技术,以‘一尺瓜’番木瓜品种为材料,通过对田间成年结果树截干后萌生的侧芽进行消毒处理,经过初代培养和增殖培养后获得大量丛生分化芽。以健壮分化芽为材料横向切取1~2.5 mm厚度的薄层6~10片,经不定芽诱导和增殖培养后进行生根培养,成功获得植株再生。研究结果表明,采用300 mg/L利福平溶液浸泡并在摇床上140 r/min振荡处理1 h、75%酒精处理30 s、0.1%的氯化汞溶液(W/V)处理7~8 min的组合递进式消毒处理方法,可使侧芽接种培养成功率达到85%以上。分化芽薄层在MS培养基+30 mg/L蔗糖+0.1 mg/L6-BA+0.2 mg/L NAA+0.3 mg/L KT+0.1 mg/L IBA的条件下诱导培养20 d,单个分化芽切取的薄层平均可获得7.04个不定芽,其中第3号薄片出芽能力最强,平均出芽数可达到3.4个,最高5.0个。采用将叶尖代替分化芽茎基部插入培养基的茎段悬空植叶促根方法进行分化芽的促根培养,生根培养基组成为1/2MS培养基+3%蔗糖(W/V)+2 mg/L IBA,20~25 d后平均出根率可达到87.5%,根系无结构疏松和愈伤化现象,假植成活率可高达95%以上,有效改善了常规生根技术存在的根系质量差和假植成活率低的问题。本研究结果可提高番木瓜组培快繁效率,并为番木瓜诱变育种和分子育种提供技术支撑。     

A tissue culture technique for vegetative propagation and low temperature preservation of Beta vulgaris

Summary A tissue culture technique for clonal propagation of Beta vulgaris is described. Flower buds or flower bud clusters formed adventitious shoots on half-strength Murashige & Skoog's medium supplemented with 10 mol/l benzyladenine (BA). The shoots were multiplied by axillary bud proliferation on a medium with 1 mol/l BA and rooted on a medium with 10 mol/l indolebutyric acid. The plants were vegetative. No mutations were observed. Genotypic variation was found in shoot formation, shoot multiplication and rooting. Some genotypes showed leaf malformations which were attributed to BA. Rooted plants in culture tubes could be stored for one year at 2 or 5°C and a low light intensity.     

Efficient Micropropagation Protocol for <Emphasis Type="Italic">Jatropha</Emphasis> Curcas Using Liquid Culture Medium

Although several studies have been made on the micropropagation of Jatropha curcas using agar base mediums, none of them have been by using liquid medium systems. The effects of explant type and temporary immersion system (test tube, jar with filter paper boat, and growtek bioreactor) on the micropropagation of J. curcas were studied. The explant type influenced shoot quality, multiplication coefficient (MC), and rooting. Leaf explant produced more and longer shoots than nodal explant. Use of filter paper (FB) boat prevented hyperhydricity and allowed proliferation of nodal explants cultured in liquid MS (Murashige and Skoog) medium supplemented 6-benzylaminopurine (BAP) and Kinetin (KN). The best shoot bud induction (92.1±3.1%) was achieved in liquid MS medium supplemented with 2.0 mg/L KN. Leaf regeneration efficiency was compared in growtek bioreactor and in jar containing liquid MS medium supplemented with 0.5 mg/L Thidiazuron (TDZ). The best shoot bud regeneration (78.7±2.1%) was obtained in growtek bioreactor. Shoot buds achieved from nodal segment and leaf were subcultured on filter paper boats in jar and bioreactor containing liquid MS medium supplemented with BAP, Indole butyric acid (IBA), Indole-3-acetic acid (IAA), and KN. Best shoot proliferation and elongation was obtained in filter paper boats containing liquid MS medium supplemented with 1.5 mg/L BAP, 0.5 mg/L IAA, and 0.2 mg/L KN. The number of multiple shoot buds was higher in leaf explants as compared to nodal explants and the highest number of multiple shoot buds was recorded from leaf explants. Up to 76.4% rooting efficiency was obtained when the shoots were ex vitro rooted. The generated plants well established in the nursery and grew normally in outdoor conditions. The protocol has good potential for application in large-scale propagation of J. curcas using liquid medium.     

Efficient In Vitro Shoot-regeneration Systems in Cassava (Manihot esculenta Crantz)

Two different protocols for in vitro regeneration of cassava using zygotic embryos and nodal axillary meristems have been developed. In both cases, buds were regenerated directly from excised explants without an intervening callus phase after a two-step culture procedure. In cotyledonary explants derived from zygotic embryos, prolific shoot formation occurred within 2—3 weeks on MS medium supplemented with 0.5—5 mg/1 BAP alone or in combination with 0.1 mg/1 NAA. Nodal explants with axillary meristems derived from aseptically grown seedlings or stem cuttings were used to initiate a round compact bulb-like structure on MS medium containing 10 mg/1 BAP. These latter structures, when cultured on MS medium supplemented with 0.1 mg/1 NAA, 1 mg/1 BAP and 0.1 mg/1 GA₃, produced multiple shoots. Somatic embryos isolated at the globular/torpedo stage from zygotic embryo explants were also capable of multiple shoot production on medium with 1 mg/1 BAP. Rooting of regenerated shoots exceeded 95 % in phytohormone-free MS medium. No change in their ploidy levels was observed. Therefore, the protocols developed should be of use in the particle gun and Agrobacterium-mediated genetic transformation of cassava.     

茄子花药愈伤诱导及不定芽分化研究

以4个茄子品种的花药为外植体,研究茄子诱导花药愈伤组织及不定芽分化.结果表明,西安绿茄,豫艺2号主要分化黄白色紧密型愈伤,这种类型的愈伤分化不定芽较多,翠绿二号,黑仙子分化黄白色松散型愈伤.松散型和紧密型愈伤组织均能分化出绿苗,但松散型愈伤分化的绿苗畸形率高,容易生长缓慢,玻璃化.豫艺2号和西安绿茄适合在F1培养基上分...     

“渝苏303”甘薯离体形态发生过程中生理生化特性的变化

甘薯茎段、叶片和叶柄在附加有1.0 mg/L NAA的MS培养基中培养5天可产生大量的不定根,培养20 d茎段和叶片分化出不定芽。本文以甘薯品种“渝苏303”为材料,研究了甘薯外植体在不定根和不定芽分化过程中可溶性蛋白质含量、过氧化物酶（POD）和超氧化物酶（SOD）活性的变化,以及可溶性蛋白质和SOD同工酶酶谱的变化。结果表明     

薄麻壳泡核桃芽接处理方式对成活率和生长的影响

为扩充薄麻壳泡核桃优株无形性繁殖材料,采用其优株穗条进行芽接,设置不同部位芽、不同嫁接高度和不同嫁接时间处理对其萌动、生长和嫁接成活率的影响,筛选出最佳芽接处理方式,提高芽接成活率.结果表明,优株对新梢长度的主效应影响显著,对芽萌动平均天数、新梢粗度和嫁接成活率的主效应影响不显著.嫁接时间对芽萌动天数、新梢长度、新梢粗...     

In vitro adventitious bud techniques for vegetative propagation and mutation breeding of potato (Solanum tuberosum L.). II. Significance for mutation breeding

Summary It was investigated what potentialities for mutation breeding of potato are offered by using adventitious sprouts that arise in vitro from leaf explants (rachis, petiole, leaflet-disc) after X-irradiation. Mutation frequency and chimerism were studied in subterranean and aerial parts in three vegetative generations (vM₁, vM₂, vM₃). Plants obtained from irradiated series produced a very high mutation frequency, a wide mutation spectrum and a very low rate of chimerism. Mutations were observed also in control series, especially in plants derived from the rachis and petiole explant group.