RAV基因对拟南芥和大豆不定芽再生的影响

RAV基因是具有AP2/ERF结构域的转录因子家族成员之一,以拟南芥rav突变体和GmRAV干涉(GmRAV-i)大豆的不同外植体为材料进行愈伤组织诱导,分别在芽诱导培养基中加入不同浓度的外源细胞分裂素2-ip和6-BA,考察平均不定芽数和不定芽再生频率。结果表明:与野生型材料相比,拟南芥rav突变体和GmRAV-i大豆在适宜的2-ip和6-BA浓度下平均不定芽数和不定芽再生频率均有所降低。证明RAV基因同系物在不同植物间具有功能保守性,并且在细胞分裂素存在下促进植物不定芽的再生。     

2个葡萄品系外植体愈伤组织诱导和植株再生

进行了无核白、红地球2个葡萄品系外植体愈伤组织诱导和植株再生的研究。通过葡萄品系无核白、红地球叶片和叶柄外植体的愈伤组织的诱导、再分化,以器官发生途径实现植株再生。结果表明:(1)愈伤组织诱导以MS+BA5mg/L+NAA0.5mg/L+蔗糖40g/L培养基最佳,诱导率为35%;同一葡萄品种的叶片与叶柄诱导愈伤组织的频率基本相同,而红地球外植体诱导愈伤组织的频率高于无核白;(2)不定芽的诱导再生以1/2MS+BA0.5mg/L+NAA0.05mg/L+CH500mg/L+蔗糖30g/L培养基最佳,平均诱导率达35.4%;叶柄来源的愈伤组织诱导不定芽的频率高于叶片愈伤组织;(3)根系的诱导以1/2MS+KT0.5mg/L+IBA1.0mg/L+AC2g/L+蔗糖15g/L培养基最佳,无核白和红地球来源的不定芽诱导生根频率基本相同,都在80%以上。     

桂竹香花器离体培养的研究

以桂竹香花柄，花瓣，花丝为外植体，研究不同激素对诱导愈伤组织和不定芽的影响，研究结果表明，4种外植体均能形成愈伤组织，但愈伤组织的发生率，发生部位及形态等表现差异，花柄愈伤组织继代MS＋2，4－D＋BA培养基，可分化产生不定芽，出芽率78％－82％，再生小苗在1／2MS＋NAA 0．5mg/L培养基中生根形成再生植株。     

二步培养和AgNO3对甘蓝型油菜子叶和下胚轴芽再生的影响

以甘蓝型油菜（Brassica napus L.）品种“浙双758”为材料,研究二步培养及添加AgNO3对甘蓝型油菜子叶和下胚轴外植体离体再生的影响。外植体先在含0.5-1.5 mg/L 2,4-D的MS培养基上 “预培养”3 d 或7 d诱导愈伤组织的产生,再转到含有3 mg/L BA和0.15mg/L NAA及添加或不添加2.5 mg/L AgNO3的分化培养基上诱导芽的分化。结果表明,外植体愈伤组织诱导率和芽再生率与2,4-D浓度、预培养时间和AgNO3密切相关;分化培养基中添加银离子可显著增加不定芽的再生频率;二步培养及添加AgNO3可使半子叶、完整子叶和下胚轴外植体芽再生频率分别达到了96.1%,96.7%和96.7%。     

花生遗传转化高效基因型的筛选与研究

选用16个花生栽培品种（系）的胚小叶作外植体,经农杆菌介导途径进行遗传转化,研究基因型差异对花生愈伤组织诱导率、不定芽分化率以及基因转化效率的影响。结果表明,不同基因型花生愈伤组织诱导率、不定芽分化率以及基因转化率差异显著。鲁花12号和02P181的愈伤组织诱导率分别为96.23%和95.59%,显著高于其他品种,P03-4和D165的愈伤组织诱导率分别为67.46%和67.33%,显著低于其他品种。05A110的不定芽分化率最高,为71.69%,其次是花选9号,不定芽分化率为46.51%,莒南2号和临花5号的不定芽分化率分别是43.71%和42.69%,与花选9号差异不显著,不定芽分化率较低的品种是E1、P03-4、远育16-8、抗青19号和潍9816,D165未分化出不定芽。临花5号的基因转化率最高,为3.36%,其次是莒南2号其转化率为1.99%,HY-1转化率为1.57%,05A106、潍9816、DS-1和花选9号四品种的转化率在0.13%-0.18%之间,未获得其他9个品种的转基因植株。     

印楝愈伤组织培养和植株再生

用印楝幼嫩枝条和叶片,在含有ＢＡ（０￣０．５ｍｇ／Ｌ）和ＮＡＡ（０￣２ｍｇ／Ｌ）的ＭＳ培养基中,于２５℃±２℃下暗培养１５￣２５ｄ,可诱导产生愈伤组织。愈伤组织生长出现２个生长峰,愈伤组织生长期和培养基中激素组成影响双峰产生时间,但不愈改变愈伤组织生长的双峰性质。在诱导不定芽培养基中,于２５℃±２℃下光培养１５￣３０ｄ,愈伤组织产生不定芽分化。愈伤组织不定芽分化与愈伤组织来源和不定芽诱导培养基的激     

甜菜组织培养与植株再生体系的建立

本研究以甜菜品种"甘糖7号"叶柄为外植体,通过诱导愈伤组织、愈伤组织分化不定芽、芽生根和移栽等步骤建立一套甜菜组织培养与植株再生体系。结果表明,诱导甜菜叶柄愈伤组织的最佳培养基为MS+4 mg/L 6-BA+0.5 mg/L NAA,诱导率达88.2%;诱导愈伤组织分化不定芽的最佳培养基为MS+4 mg/L 6-BA+0.5 mg/L NAA,分化率仅为9.4%;生根培养基为MS+0.1 mg/L NAA,生根率达33.3%。甜菜再生苗移栽至蛭石中,保温保湿7~10 d后,成活率可达90%以上。本研究结果将对糖料作物甜菜抗逆遗传改良及其基因工程育种具有推动作用。     

滁州白头翁组织培养及再生体系的建立

为建立白头翁再生体系,以白头翁主根的切段和试管苗叶片为外植体,比较2种外植体离体培养差异,探讨不同植物生长调节剂对不定芽和愈伤组织诱导、愈伤增殖和再分化及芽苗生根的影响。结果表明:叶片可以通过愈伤组织再分化和直接产生不定芽2种途径建立再生体系,而根只诱导出了愈伤组织,增殖培养中逐渐褐化死亡;在含6-BA培养基上,叶块死亡,而根只诱导出少量愈伤组织;叶片诱导不定芽的培养基为MS+TDZ 0.3 mg/L+NAA 0.1 mg/L;愈伤组织增殖的培养基为MS+TDZ 0.2 mg/L+2,4-D 0.2 mg/L;再分化时,需要转接到TDZ和NAA组合的培养基上,其中处理组合TDZ 0.3 mg/L+NAA0.1 mg/L的再分化率达100%;生根培养基为1/2MS+NAA 0.2 mg/L+IBA 0.2 mg/L+蔗糖20 g/L。不同外植体离体培养存在差异,叶片较根易培养;TDZ对白头翁叶片培养效果较好,2,4-D对愈伤组织的诱导能力较强,而NAA适合于不定芽分化,NAA与IBA组合使用生根效果较好。     

平贝母愈伤组织和不定芽诱导研究

目的应用组织培养技术对平贝母进行离体培养,从而得到愈伤组织和不定芽。方法以鳞茎为材料诱导愈伤组织与不定芽,采用正交实验设计确定最佳的激素种类与浓度。结果适当的激素组合及配比可诱导出愈伤组织和不定芽,最高的诱导率可分别达到14%以上、86.7%。结论愈伤组织最佳诱导培养基为MS+2,4-D 1.0mg/L+6-BA 0.9mg/L+NAA 0.5mg/L;最佳继代培养基为MS+2,4-D0.5mg/L+6-BA 0.3mg/L;不定芽最佳诱导培养基为MS+KT 0.8mg/L+NAA 0.3mg/L。     

普通大麦(Hordum vulgare)未成熟胚愈伤组织的诱导和植株再生的初步研究

本文对五种基因型的普通大麦未成熟胚在离体培养下愈伤组织的诱导能力和植株再生能力进行了比较。初步结果表明:0.5—1.5毫米大小的未成熟胚为合适的培养材料,只要诱导培养基合适,各基因型都能诱导出愈伤组织,并达到较高的诱导率。但愈伤组织的质量在各基因型中存在显著差异。同时发现,再生植株诱导频率主要决定于愈伤组织的质量,而愈伤组织的质量直接由基因型和诱导培养基决定。不同的分化培养基在分化再生植株的作用上没有出现显著的差异。     

东北春玉米骨干自交系组织培养再生系统研究

东北玉米骨干自交系在诱导愈伤组织和植株再生中,基因型差异非常显著。自交系7922最高愈伤诱导率达89.1%,芽苗率为66.4%;而最低的921诱导率仅有18.9%,芽苗率为0。再生途径主要有两种:7922主要以体胚发生途径为主,4122、8902主要以器官发生途径为主。2熏4-D是体胚诱导不可缺少的植物生长激素,适当的细胞分裂素能促进诱导4112、8902不定芽的产生。而且,再生途径并不是一成不变的,在一定条件下可以相互转变。     

In vitro shoot regeneration ofSolanum tuberosum cultivars: interactions of medium composition and leaf, leaflet, and explant position

Summary Shoot regeneration was investigated on explants from different leaves and leaflets of three potato cultivars Posmo, Folva and Oleva. Explants were excised from glasshouse grown plants and grown for 6 days on callus induction medium with indole-3-acetic acid or 2,4-dichlorophenoxyacetic acid. Explants were then transferred to auxin free shoot regeneration medium with gibberellic acid and 6-benzyladenine or zeatin. By using the optimum combinations and concentrations of plant growth regulators and by excision of explants from particular regions of proximal leaflets from newly unfolded leaves, shoot regeneration frequencies of 97.0% were obtained for cv. Posmo and 32.1% for cv. Folva. Shoot regeneration frequency of cv. Oleva was very low and could not be improved by the different treatments.     

Callus induction and in vitro plant regeneration of rice (Oryza sativa L.) under various conditions

The objective of the present study was to develop an effective protocol for optimum callus induction and complete plant regeneration for four varieties of rice (Oryza sativa L.) i.e., Super Basmati, Basmati-370, Basmati-371 and Fakhre Malakand. Calli were induced from mature seed scutelum. The Murashige and Skoog (MS) and Chu's N6 media containing hormone 2, 4-D (2, 4-Dichlorophenoxy acetic acid) in different concentrations were used for callus induction. Fakhre Malakand produced maximum calli on N6 media containing 3 mg L(-1) 2,4-D. while other three varieties showed maximum callus induction on N6 media containing 2.5 mg L(-1) 2,4-D. N6 media was found better than MS media for callus induction. For complete plant regeneration the calli of two varieties i.e., Basmati-370 and Basmati-371 were plated on N6 media containing different concentrations of NAA (1-Naphthalene acetic acid) and BAP (6-benzyl aminopurine). The maximum regeneration frequency (%) was observed on N6 media containing NAA 1 mg L(-1) and BAP 2.5 mg L(-1). It took 27-30 days for the callus to regenerate into a complete plant. Basmati-370 produced 4-7 plantlets per callus whereas Basmati-371 produced 4-8 plantlets per callus with regeneration frequencies of 61 and 69%, respectively.     

Regeneration ofSolanum tuberosum cv. Katahdin from leaf expiantsin vitro

A previously published medium has been found to be effective for plant regeneration from leaf expiants ofSolanum tuberosum cv. Katahdin. A number of combinations of thidiazuron and naphthaleneacetic acid, along with a medium previously published for shoot regeneration from protoplast-derived calli ofSolanum phureja, which contained zeatin, indoleacetic acid and gibberellic acid (ZIG), were tested for callus induction, and all calli were transferred to ZIG for shoot induction. We conclude that leaf expiants cultured on ZIG medium at all stages of culture were the most effective at shoot production. A mean of over six shoots were produced per expiant after four months in culture.     

第一例栽培种花生花药培养单倍体植株的创制

以栽培种花生为材料,开展花生单倍体培养的前沿性探索,包括离体花药诱导形成愈伤组织、愈伤组织分化形成幼芽、再生苗培养、再生植株倍性鉴定和嫁接移植等研究。通过比较外植体灭菌时间、诱导培养基、分化培养基、再生培养基和再生植株创制培养条件等试验,筛选出适合花生花药培养的方法：1%NaClO消毒灭菌9 min,愈伤组织的诱导培养采用B5N1、B3N1培养基,再分化培养采用B5N1-2培养基,再生苗培养采用SG培养基。本研究获得1株花药培养的再生植株,编号为15B8-8。荧光原位杂交技术（FISH）鉴定结果表明,该植株具有20条染色体,其中9条为A染色体、11条为B染色体,是第一例来自栽培种花生花药培养的单倍体植株;研究还表明,汕油52花生品种具有较强的花药培养力,有潜力作为花生花药培养的“桥梁品种”。     

紫罗兰愈伤组织诱导及植株再生的研究

以紫罗兰幼苗子叶、子叶柄、下胚轴作外植体接种MS附加不同激素的培养基诱导愈伤组织，并进一步诱导分化出芽及再生植株。子叶和下胚轴外植体在MS＋0.1mg/LNAA培养基上愈伤组织发生率达100％，下胚轴愈伤组织转移MS 0.1mg/L 6-BA培养基上易诱导分化出小苗，分化频率达67％，再生小苗在1／2MSA＋0.2mg/L IBA培养基上生根率达86％。     

Plant regeneration from leaf tissues of four North Dakota genotypes of potato (Solanum tuberosum L.)

An efficient plant regeneration system was developed fromin vitro leaf tissues of four North Dakota potato genotypes. The best medium for genotype ND860-2 was Murashige and Skoog medium with 20.0 μM IAA and 11.4 μM zeatin riboside. Two to three weeks of initial dark treatments had a significant effect in reducing the time for shoot regeneration and increasing the number of regenerated shoots. Four antibiotics commonly used inAgrobacterium- mediated transformation were tested for their effects on shoot regeneration. Shoot induction was completely inhibited by kanamycin at 15 mg/L and hygromycin at 4 mg/L or higher, but not by carbenicillin (except at 1000 mg/L) and cefotaxime. Hygromycin significantly stimulated shoot induction at 1 mg/L.     

不同基因型菜心游离小孢子培养和植株再生

以11个不同基因型的菜心栽培品种为试材,研究不同培养条件对菜心游离小孢子胚诱导和植株再生的影响。结果有7个基因型材料获得小孢子胚,从不同基因型诱导形成胚的频率存在显著差异,表明基因型是影响菜心小孢子胚发生的主要因素;第1天热激培养时用170 g/L高浓度蔗糖培养之后转换成含130 g/L蔗糖培养基能显著提高小孢子胚诱导率;0.05 mg/L 6-BA+0.2 mg/L NAA能促进菜心小孢子诱导成胚;添加0.4 mg/L GA3可显著提高菜心小孢子胚芽诱导率和平均每胚出芽数。7个基因型材料均诱导出再生植株,植株诱导率为100%。     

神农香菊丛生芽的诱导及植株再生

以神农香菊茎段为外植体,研究植物生长调节剂及活性炭(AC)对神农香菊丛生芽诱导的影响,以建立神农香菊丛生芽诱导及植株再生的高频再生体系。结果表明,丛生芽诱导的最适培养基为MS+2.0 mg/L 6-BA+0.05 mg/L NAA,最适增殖培养基为MS+2.0 mg/L 6-BA+0.05 mg/L NAA+0.05 g/L AC,增殖系数为45.3,添加AC可有效促进玻璃化苗恢复;生根诱导的最适培养基为不添加任何生长调节剂的1/2MS,生根率高达100%;将生长良好的再生植株进行移栽,存活率可达100%。     

紫色甘薯的茎尖培养与脱毒

以紫色甘薯品系为试材,研究6-BA和NAA及基因型对紫色甘薯茎尖培养的影响,并采用硝酸纤维素膜—酶联免疫法(NCM-ELISA)和症状法对再生植株进行病毒学检测。结果表明,培养基中6-BA和NAA浓度和配比对紫色甘薯茎尖培养的愈伤组织、不定根和不定芽的诱导有明显影响。6-BA可促进不定芽的诱导,附加1mg/L6-BA的MS培养基为最佳诱芽培养基。基因型对诱芽率也有一定的影响,其中品系B3和B7的诱芽率可达50%。通过病毒学检测,获得12个紫色甘薯品系的脱毒苗,其平均脱毒率为94.7%。