青花菜小孢子再生植株加倍及倍性鉴定

采用秋水仙碱溶液对单倍体植株进行诱导,并采用流式细胞仪及保卫细胞叶绿体染色法对青花菜小孢子再生植株群体的倍性进行分析。结果表明,小孢子再生植株群体单倍体和二倍体所占比例基本一致;采用200 mg/L秋水仙碱处理单倍体植株20 h后,3个供试基因型植株的存活率分别为88.9%、90%和100%,二倍体诱导率分别为66.7%、70%和77.8%,另外还得到1株四倍体植株。采用FDA荧光染色和1%I-KI染色法可清楚地观察到不同倍性间植株气孔大小和形态以及保卫细胞叶绿体数存在显著差异;单倍体植株气孔长度约为20μm,二倍体约为25μm,而四倍体大于33μm;单倍体保卫细胞叶绿体数为7个左右,二倍体约13个,而四倍体最多超过23个。1%I-KI染色法对设备要求简单,宜普遍采用。     

辐照花粉诱导西葫芦单倍体

摘　要：利用不同剂量的60Co γ射线辐照西葫芦花粉后授粉,剥取不同发育阶段的胚接种在E20A培养基上进行培养,研究不 同基因型、辐照剂量、剂量率对西葫芦单倍体诱导的影响。结果发现：单倍体的产生和植株再生率受基因型、胚发育阶段和 辐照剂量的影响明显。281基因型平均单倍体诱导率最高,为1.30 %;点状胚和箭形胚易发育成单倍体苗;辐照剂量80~100 Gy时获得点状胚和箭形胚及胚总数最多,杆状胚的植株再生率和植株成活率最高,分别是83.33 %和35.71 %;适宜的剂量率 为45.19 Gy·min^-1。     

辐射花粉授粉诱导甜瓜单倍体

为了掌握辐射花粉授粉诱导甜瓜单倍体技术,以5种基因型甜瓜为材料,通过应用γ射线辐射的花粉进行授粉结合胚胎培养,从2种基因型中获得了单倍体植株,单倍体的平均诱导频率为0.29%。这2种基因型分别属于厚皮和薄皮甜瓜类型。花粉的辐射剂量为300Gy和600Gy时,供试材料的平均坐果率分别为50%和10%。辐射剂量为300Gy时,诱导厚皮甜瓜4810和薄皮甜瓜新富玉形成了单倍体植株,单倍体植株的诱导频率分别为0.55%和0.63%;剂量为600Gy时未能诱导供试材料形成单倍体。研究结果表明母本的基因型和花粉的辐射剂量对坐果率和单倍体的产生有一定的影响。此外,对单倍体植株与正常二倍体植株的形态学进行了比较观察,结果表明单倍体植株生长势弱,叶片较小,雄花和雌花均败育。     

紫外辐照花粉授粉诱导西瓜单倍体研究

以6种不同基因型的西瓜为材料,通过用紫外线辐射的花粉授粉结合胚胎培养,研究不同基因型、辐照剂量和胚挽救时间对西瓜单倍体诱导的影响,并对再生苗进行倍性鉴定。此外,还对诱导培养基和生根培养基进行优化。结果表明:单倍体的诱导率和植株再生率受基因型、胚挽救时间和辐照剂量的影响,而且还与季节有关。     

胡萝卜游离小孢子培养与植株再生技术研究

对"长城红玥"、"宝冠"2个胡萝卜自交品种进行游离小孢子培养,研究花序与小孢子发育的相关性,以期获取游离小孢子和最佳灭菌的方法。结果表明:不同基因型间游离小孢子培养难易存在差别,通过诱导胚性愈伤组织,芽分化,成功培养出单倍体和双单倍体再生植株,再生株系46.9%植株是单倍体,53.1%植株是双单倍体,诱导胚性愈伤组织、分化再生植株和生根的最佳培养基分别是HLB+0.5 mg/L 2,4-D、MS+0.5 mg/L 6-BA和MS(或W14)+1.0mg/L NAA。     

西瓜未授粉子房的离体培养

为了探讨西瓜未授粉子房诱导率的影响因素,以‘早春红玉’、‘西农9号’和‘小绿皇’为供试材料进行西瓜未授粉子房的离体培养,研究供体植株的黑暗热激处理、基因型、不同取样时间、2,4-D浓度和不同激素浓度组合等因素对其芽点诱导率的影响。结果表明:西瓜未授粉子房离体培养以33℃的条件下黑暗热激4 d效果最佳;以开花前1 d的子房诱导率最高为17.2%;3个品种中仅‘早春红玉’获得了再生植株,且在激素组合为4.0 mg·L-12,4-D+2.0 mg·L-16-BA+0.5 mg·L-1NAA时芽点率最高为15.0%;根据再生植株的根尖染色体数目,初步鉴定再生植株中有单倍体植株,还有二倍体和四倍体植株。     

‘嘎拉’苹果小孢子来源纯合基因型植株叶片再生体系研究

【目的】建立‘嘎拉’苹果小孢子来源纯合基因型植株高效再生体系,探索一个简单有效的苹果纯合基因型株系叶片外植体诱导再生不定芽的方法。【方法】以‘嘎拉’苹果花药离体培养获得的纯合基因型株系离体叶片为外植体,进行再生培养,检测‘嘎拉’各纯合基因型株系的不定芽再生能力;通过优化植物生长调节剂、叶片预处理和培养方式,建立苹果纯合基因型植株高效再生体系。【结果】苹果纯合基因型株系中双单倍体株系DH2-10的再生率最高,四倍体纯系DH2-15次之,单倍体株系DH2-3再生率较低,三倍体纯系DH2-40在培养中难以再生。不同基因型适宜的激素质量浓度不同,相较于双单倍体株系,四倍体纯系DH2-15在较高细胞分裂素水平再生率较好。再生培养过程中,整叶带伤口的外植体预处理方式优于叶片切块接种的方式,接种苗龄40 d左右的叶片有更好的再生率。【结论】建立了苹果纯合基因型高频高效离体再生体系:双单倍体株系DH2-10在最适再生培养基MS+0.5 mg·L~(-1)IBA+5 mg·L~(-1)6-BA上培养,再生周期为23~35 d,再生系数为7.27,再生率达100%。四倍体纯合基因型株系DH2-15适宜在MS+0.5 mg·L~(-1)IBA+6 mg·L~(-1)6-BA培养基上进行再生培养,再生率为93.33%。建立的再生体系可用于纯合基因型株系的快速增殖和遗传转化。     

马铃薯四倍体栽培品种花药培养及再生植株的鉴定

以马铃薯四倍体栽培品种讷16、波C、克新13号和扎列娃为试材进行花药培养,研究不同基因型花药培养的效率及再生植株的鉴定。结果表明：花蕾外部形态与小孢子的发育阶段呈对应关系,可据此确定取材时期;讷16的愈伤组织诱导率和分化率最高,分别达到27.60 %和6.04 %;双单倍体植株的生长势、叶色、薯形等性状与亲本差异明显。     

黄瓜未授粉子房诱导出胚和再生植株的研究

以7个不同黄瓜基因型为材料进行黄瓜未授粉子房的培养研究,结果表明,不同基因型出胚率差异显著,其中密刺型黄瓜北京102出胚率最高,达25.3%,鄂黄3号出胚率最低,为0.7%。以北京102、中农26号和津优1号为试材进一步研究热激诱导、外源激素、植株不同生长阶段对胚状体诱导产量的影响,发现35℃热激处理2d胚状体产量最高,分化培养基中NAA为0.20mg·L-1、6-BA为0.8mg·L-1时最易分化出胚,在植株第21~30节位采收的子房出胚量最高。再生培养基中GA浓度达到0.4~0.8mg·L-1时,可有效打破再生植株"花打顶"现象从而提高植株再生率。再生植株中二倍体植株比率为70.0%,二倍体植株中DH株比率为84%。     

黄瓜(Cucumis sativusL.)花药培养

以卡罗尔、HH1-8-57与津绿农家乐3个不同基因型黄瓜品种为试材,在Kumar等、Song等和詹艳等试验研究基础上,设计试验分别探讨低温预处理时间、培养基成分(胚性愈伤组织诱导培养基、胚状体诱导培养基)和基因型等因素对黄瓜花药培养的影响。结果表明:在4℃低温预处理2d时,胚性愈伤组织诱导率显著高于其他处理;在MS+1.0mg·L-12,4-D+0.5mg·L-16-BA+3%蔗糖+0.8%琼脂培养基上胚性愈伤组织诱导率最高,达到81.3%;在MS+0.1mg·L-1NAA+3.0mg·L-16-BA+3%蔗糖+0.8%琼脂培养基上胚状体诱导率最高,为40.0%;基因型间胚性愈伤组织诱导率及胚状体诱导率差异显著,津绿农家乐品种胚性愈伤组织诱导率最高,达到81.1%,HH1-8-57胚状体诱导率最高,为40.0%。本研究从2份试材中成功地诱导出胚状体并获得了黄瓜单倍体再生植株。     

Physiological alterations modulated by rootstock and scion combination in cashew under salinity

In this study we evaluated the influence of rootstocks and scions on physiological disturbances that are induced by salinity in cashew (Anacardium occidentale) plantlets. Two cashew genotypes, CCP 09 and BRS 226, were utilized as rootstocks and scions, resulting in four scion/rootstock combinations by reciprocal grafting. The plantlets were irrigated in absence (control) or in presence of 50 or 200 mM NaCl for 15 days under greenhouse conditions. The plantlets with BRS 226 as rootstocks demonstrated higher transpiration and greater accumulations of Na⁺ and Cl⁻, proline and free amino acids in leaves compared to plantlets that having CCP 09 as rootstocks. The K⁺ content in roots and leaves of all four combinations was not influenced by salinity or by different scion/rootstock combinations. The self-grafting of the BRS 226 genotype showed the highest stability for chlorophyll and Rubisco, exhibiting the highest tolerance to salinity. The scion genotype did not affect any of the studied physiological parameters. The studied physiological disturbances induced by salinity in cashew plantlets were more influenced by rootstock than by scion and these changes were also dependent on compatibility between scion and rootstock.     

辣椒花药培养影响因素的研究

以6种基因型辣椒的花蕾为材料,比较了不同基因型辣椒花蕾的消毒效果,辣椒基因型及花药接种量对花药培养的影响。研究结果表明,本试验的消毒方法获得了较好的消毒效果,3个基因型材料均未受污染,其他最高的污染率为23.08%;50%的基因型能诱导出胚状体,出胚率1.18%～1.38%,小果型的辣椒不能诱导出胚状体;所有基因型均能诱导出愈伤组织,出愈率17.65%～69.44%;花药接种量对花药愈伤组织诱导有影响,增大接种量,出愈率上升0.77%～13.67%,但对胚状体诱导影响不明显;在愈伤组织的继代培养中,接种量对其生长的影响因基因型不同而不同,所有基因型的愈伤组织在第一次继代培养中均不能分化出胚状体或不定芽。     

Rapid in vitro multiplication and conservation of Garcinia indica: A tropical medicinal tree species

A simple and efficient method has been developed for rapid regeneration of plantlets via adventitious bud differentiation on mature seeds of Garcinia indica (Thouars) Choisy, a medicinally important facultative apomictic tropical tree species. High frequency direct shoot proliferation was induced in seed segments cultured on Murashige and Skoog's medium supplemented with cytokinins (BAP, kinetin and TDZ) alone and in combination with auxin (NAA). Amongst the various combinations used, BAP proved to be the most effective. Multiple shoots formed within 4–5 weeks of culture. The shoot forming capacity of the seeds was influenced by the BAP concentration tested (5–50 μM) and optimal response was observed at different concentrations (12.5–50 μM) in different genotypes investigated. Significant differences were recorded in terms of percent response (27.78–100%) as well as average number of shoots per explant (3.49–57.67) among the four genotypes investigated. Elongation of the induced shoots was achieved on MS basal medium containing 0.2% activated charcoal. The induction medium had a profound effect on rate of bud elongation with shoots induced on lower concentrations of BAP showing as much as four-fold elongation within 4 weeks. Proliferating shoot cultures were established by repeatedly subculturing the shoot nodes on MS medium supplemented with 5 μM BAP. Maximum rooting (91.66%) occurred in shoots cultured on half-strength MS medium supplemented with 10 μM IBA. The plantlets were successfully transferred to soil after hardening; the time period from initiation of shoot buds to transplantation being 15–17 weeks. For in vitro conservation, the shoot cultures were maintained on medium supplemented with 0.5 μM BAP and the subculture duration could be enhanced up to maximum of 11 months.     

君子兰种子离体培养的研究

试验对君子兰品种'油匠'、'胜利'、'和尚'的种子离体培养进行了研究,结果表明,直接从果实中剥离而采用70%酒精10 s 0.1%升汞8 min的消毒效果最好,污染率仅5.68%.基因型对种子组织培养的影响较大,'油匠'的种子较易于诱导出愈伤组织,诱导率可达72.97%.'胜利'的种子最适诱导培养基为MS 2,4-D 2 mg/L BA 2 mg/L,其诱导率为52.94%,最适分化培养基为MS NAA 1 mg/L BA 1 mg/L,分化率达72.22%.君子兰种子在培养基上诱导产生愈伤组织并能大量分化成苗,对君子兰组培工厂化生产有较大意义.     

苹果不同继代次数的茎尖组培苗同工酶酶谱及RAPD分析

 分别对不同继代次数(3～90代) ‘富士’、‘金冠’、‘乔纳金’苹果组培苗进行POX、EST、AMY同工酶分析。结果表明, 同一品种不同继代次数处理间均有一致的酶带谱型。利用超薄层等电聚焦电泳根据POX酶蛋白pI的差异可区分金冠和乔纳金两品种, 同一品种不同继代次数的组培苗之间pI相同,组培过程未对POX酶蛋白pI产生影响。利用25个随机引物对不同继代次数的苹果组培继代苗进行了RAPD分析, 其特征带型表现一致, 未发现变异。说明苹果茎尖离体继代培养90次以内具有较高的遗传稳定性。     

试管芋诱导的研究

 以芋组培苗为试材, 研究了蔗糖浓度、激素浓度、光照时间、培养温度、不同大小试管苗,不同芋品种类型等因素对试管芋诱导的影响及不同基质对试管芋育苗成活率的影响。结果表明: 诱导试管芋较理想的培养基为MS + 蔗糖8 % + BA 1. 0 mg·L ^{- 1} + NAA 0. 5 mg·L^-1 , 光照12 h/ d , 培养温度30 ℃; 试管苗越大越有利于试管芋的形成; 试管芋可保持其原有的特性; 基质影响试管芋育苗的成活率, 4 种基质中蛭石最好。     

利用RAPD标记研究部分葡萄品种的亲缘关系

对47个葡萄品种进行RAPD分析。从100条随机引物中筛选出30个多态性引物用于DNA扩增,共扩出216条DNA带,121条具有多态性,多态性比例为56.01%。通过聚类分析结果表明,供试品种分成5大类,巨峰、红瑞宝、龙宝等巨峰系品种聚为一类。其中龙宝和红瑞宝优先聚在一起。表明葡萄品种间的遗传多样性有明显差异,即使父母本相同的品种,利用RAPD技术也可以分开。聚类图显示,品种间遗传一致度与品种的遗传背景有关,如高妻与红井川,红瑞宝与龙宝优先聚在一起;有的品种虽属同一杂交组合却未能表现最大遗传一致度,如京玉与奥古斯特,在聚类图中处于相邻的位置,说明无性繁殖的葡萄在双亲基因型杂合条件下,有性杂交通过基因重组,杂种后代也可呈现出有较大遗传差异。     

秋水仙素处理‘罗田甜柿’获得12倍体再生植株

 以‘罗田甜柿’试管苗离体叶片为材料，研究了秋水仙素浓度及处理时间对诱导染色体倍性变异的影响。结果表明：0.3％秋水仙素处理4～6 d，可诱导染色体加倍; 与对照比较, 再生变异植株的气孔孔径增大，密度降低，根尖染色体数2n=12x=180，叶片DNA含量为对照的两倍。LI、LⅡ、LⅢ细胞层均加倍的证据表明，获得的倍性变异株为同质12倍体。     

Acclimatisation to mannitol-induced water deficit in miniature rose (Rosa hybrida) plantlets cultivated in vitro under autotrophic conditions

Summary

Miniature rose plantlets at the flower development stage were grown photo-autotrophically on MS medium and subsequently exposed to water deficits of –0.23, –0.32, –0.40, or –0.67 MPa osmotic potential ( _s) for 14 d. The _s in the culture medium was raised by increasing the concentration of mannitol, which caused abnormal floral development in terms of the flowering percentage and the number of flowers per plantlet, as well as delayed flowering. In vitro flowering and the number of flowers per plantlet declined significantly when miniature rose plantlets were exposed to water deficit stress at –0.40 MPa or –0.67 MPa. Reductions in growth, pigment degradation, chlorophyll a fluorescence, and net photosynthetic rate (Pn) were greatest in plantlets exposed to a water deficit stress of –0.67 MPa. This was particularly evident in the case of Pn, with a decline of 73.7% compared to non-stressed control plantlets. In contrast, proline levels increased in plantlets under water deficit stress, as proline performs a key role as an osmoprotectant under such conditions. The flowering stage in miniature rose plantlets is particularly susceptible to water deficit stress, which suppresses the development of reproductive organs. Knowledge of the responses to water deficit stress at the reproductive stage may be applied to identify effective indices for the selection of genotypes with increased tolerance to water deficit in miniature rose breeding programmes.