棉花试管苗“出管”移栽经验点滴

一般认为,棉花再生植株移栽比较困难。有人~（［１］）用“水培法”作为试管苗“出管”到盆栽之间的过渡,以提高移栽成活率。我们根据多年在移栽方面的经验,采用两次移栽方法移栽棉花试管苗,成活率在９０％以上。在此作一交流与探讨,以供同行参考。１出管前的准备从组织培养、原生质体培养,或转基因细胞团经胚胎发生的再生小试管苗,长至５～６片叶子时,移入较大的培养瓶,放入人工气候箱中继续培养２～４周,培养温度由原来的２８℃降为（２４±１）℃。待根部有数条白色的新根长出,即可移入蛭石花盆中。２移入蛙石花盆（第一次）…     

玉米再生植株移栽的研究

玉米在农业生产和遗传学研究中占有重要地位,组织培养再生的植株可用于品种改良和遗传学研究。如何提高玉米再生植株移栽成活率,是一个十分重要的问题。有关试管苗移栽的研究,国内外已有一些报道。我们连续三年进行了玉米试管苗移栽研究。材料为生产上推广的玉米自交系黄早4,B73,单交种烟单14、鲁玉7和鲁单40。     

提高玉米花粉植株移栽成活率的研究

玉米花药培养的成功,为大量地产生单倍体提供了新的途径,所以引起了国内外遗传育种工作者的关注。但是花粉植株移栽成活率低,是当前存在的突出问题之一。据估计,国内有80％以上的花粉植株死于移栽;Green(1978)报告用玉米盾片培养再生的植株成活率也只有10～15％。看来,通过组织培养获得的玉米植株成活率普遍较低。提高试管苗移栽成活率的研究是极有意义的课题之一。     

杂交水稻体细胞再生植株的观察

改进了杂交水稻器官发生的再生植株的炼苗方法。采用两步炼苗法再将植株直接移栽入土,可使移栽成活率超过80％。观察了温室条件下成活的体细胞再生植株的生长、发育及结实情况。结果表明,再生植株的总叶片比实生植株减少,有效分蘖增加,抽穗期提早。但单株实粒数减少,种子粒形出现变异,单株之间粒重变异显著。总计对43株再生植株的粒重进行了统计分析,发现了7个粒重变异显著的单株,变异率为16.3％。起源子茎的再生植株与起源于幼穗的再生植株相比,各方面无显著差异。比较了再生植株与在同样条件下栽培的实生植株的种子发芽率,发现个别再生植株的种子发芽率有所下降。     

三叶半夏叶片一步成苗离体培养技术

以药用植物三叶半夏叶片为材料,通过比较直接和间接器官发生两种途径,建立了半夏一步成苗的快速繁殖技术体系。结果表明,经过愈伤组织阶段的一步成苗培养基为MS+0.5mg/L2,4-D+1.0mg/LKT,90d左右方可得到再生植株,植株分化率为74%,每个外植体上分化的块茎数为5.61±1.04。附加NAA与BA两种激素对一步成苗培养基进行优化,筛选出一步成苗最佳培养基MS+0.5mg/LNAA+0.5mg/LBA,60d后就可直接发育成完整植株,植株分化率为76%,每个外植体上分化的块茎数高达9.97±0·81,对这种培养基上的再生小植株进行移栽,1个月后,移栽成活率达100%。     

草莓组织培养研究初报

取草莓茎尖,进行离体培养,分化出再生植株、完正植株放于低温(5—10℃)处,壮苗锻炼、获得同步苗、移栽成活率80％以上,通过电镜检测茎尖苗无病毒粒体、无毒草莓所内小区试验获得增产13.9～22.48％的明显效果。     

西瓜组培苗嫁接技术的研究

西瓜组培苗往往不易移栽成活,且生长差。通过砧木嫁接的方法,可提高苗的成活率并改善苗的生长状况。实验研究了接穗炼苗的天数、接穗叶片数、接口用不同外源激素处理与嫁接成活率的关系。条件优化后,嫁接后成活率可达85％,为西瓜优良品种的快速繁殖以及西瓜抗虫、抗病、品质改良等基因工程的研究奠定了基础。     

6个烟草杂交组合花药再生苗的培养和DH群体的构建

以6个烟草(Nicotiana tabacum L.)杂交组合的花药为实验材料,对各杂交组合花药再生苗出苗状况及培养过程中添加H液体基本培养基对花药出苗状况的影响进行了比较分析;在此基础上,采用质量体积分数0.4%秋水仙素浸苗法构建加倍单倍体(DH)群体,对加倍处理后不同杂交组合再生苗的成苗率、大田移栽成活率及染色体加倍率等进行分析,并对杂交组合DH1单倍体和加倍单倍体植株叶片气孔保卫细胞叶绿体数的差异进行了研究。结果表明:不同杂交组合每枚花药的出苗数、加倍处理后再生苗的成苗率、大田移栽成活率及染色体加倍率有较大差异。其中,每枚花药出苗数为1.57～3.30,杂交组合DH5每枚花药的出苗数最多,达到3.30株,显著高于其他5个杂交组合(P<0.05);加倍处理后再生苗的成苗率为21.12%～33.42%,大田移栽成活率为91.87%～98.86%,杂交组合DH6的成苗率和大田移栽成活率最高;染色体加倍率为6.64%～10.78%,杂交组合DH4的染色体加倍率最高。花药培养约15 d后,添加H液体基本培养基能够显著促进杂交组合DH1和DH2花药出苗。杂交组合DH1单倍体苗和加倍单倍体苗叶片气孔保卫细胞的平均叶绿体数分别为9.27和17.46,二者比值接近1∶2,差异显著。通过花药培养和染色体加倍处理,分别获得了6个烟草杂交组合的DH群体。     

盐桦无菌材料的成功获得及其试管苗移栽定植成活的方法

目的:探讨获得盐桦无菌材料的最适灭菌方法和提高移栽定植成活率的关键性技术要点。方法:以新疆濒危植物盐桦当年生休眠枝条为材料,在适宜的培养基上筛选最适灭菌方法,继而以生长85~90d左右的组培试管苗为材料,探讨移植时期、温度、光照、水分、营养对成活率的影响。结果:采用半无菌水表面灭菌 75%酒精30s 0.1%升汞7~10min处理效果最好,污染率为13.3%。盐桦组培试管苗移栽定植中,以4月中旬~5月中旬成活率较高,平均成活率在95%以上。结论:研究筛选获得了盐桦的最适灭菌方法和盐桦组培试管苗移栽定植成活的适宜方法,为工厂化扩繁盐桦苗木创造了条件。     

沙葱(Allium mongolicum Regel)离体培养再生可育植株

以沙葱（Allium mongolicum Regel)的叶基切段作外植体,成功地诱导出愈伤组织和再生植株。诱导愈僵组织的培养基为附加2mgL^-22,4-D和0.2mgL^-1KT的MS培养基。愈伤组织转移到附加2mgL^-16BA和0.4mgL^-1NAA的MS培养基后分化出大量的苗;分化苗在含有IBA1mgL^-1的1/2MS培养基上生根最佳。再生植株移栽成活率在95%以上。移栽到田间的植株第二年以后可正常开花结实。     

Potential implications of biopriming in banana (Musa spp) plantlets against banana bunchy top virus (BBTV)

Abstract

Transplant media as a means for the introduction of biological agents is currently being investigated in a variety of crops. This study aimed to investigate the impact of microbial inoculation in micropropagated banana plantlets to enhance their resistance against Banana bunchy top virus (BBTV). Virus indexed micropropagated plantlets of banana were subjected to root colonization followed by foliar spraying with bacterial strains Pseudomonas fluorescens Pf1, CHA0 and Bacillus subtilis EPB22 during primary and secondary hardening stage in the nursery, at the time of repotting and 3 months after planting in the pot. Microbe inoculated plantlets showed enhanced PR proteins and defense enzymes besides reducing banana bunchy top disease incidence under glasshouse condition. The results indicated the effective use of beneficial microbes in reducing the disease incidence of BBTV in tissue culture banana plantlets. In addition, the molecular characterization of endophytes isolated from banana plantlets, using SDS-PAGE and RAPD-PCR revealed that endophytes were categorized into two distinct groups. These results emphasize the significance of microorganisms in protection of young plantlets from transplanting stresses in field. Further, the use of beneficial microorganisms instead of chemicals sustains an ecological niche in the agricultural ecosystem.     

An efficient grafting system for transgenic plant recovery in cotton (Gossypium hirsutum L.)

A successful transformation program relies on the number of survival plants in soil that can be obtained. Low recovery of transgenic plants is still a key restrictive factor for transgenic cotton production. In order to utilize genetic transformation in cotton breeding program effectively, an efficient grafting system for recovering plants derived from somatic embryogenesis following Agrobacterium infection and kanamycin selection was developed. Various aspects of in vitro grafting were examined in efforts to improve the efficiency of transformant recovery. Using strong seedling rootstocks was the first important step to obtain high rate of successful grafts. Scion size >0.6 cm and seedling rootstock at age of 6–12 days were appropriate for grafting. The successful grafting ratio was higher when using hypocotyls without radicle. Shoot-tip and shoot stem with axillary bud were also suitable for in vitro grafting, which meant we could significantly improve the survival ratio of transgenic plantlets, because one plantlet has a shoot-tip but several axillary buds. Based on our data, the period from in vitro seedling rootstock germination to transplant of grafts to field usually takes one month. Over 90% successful grafting ratio could be obtained under optimal conditions, which represented a significant improvement over currently available methods for recovery of cotton plantlet from somatic embryogenesis after transformation. Ex vitro grafting could also be used for plant recovery, which gave an average of successful grafting ratio of 71.9%. However, this method was strongly affected by environmental factors.     

白木香组织培养及快速繁殖

白木香腋芽外植体在含０．０５ｍｇ／ＬＢＡＰ的ＭＳ基本培养基上增殖系数为４．９３。采用间接法诱导芽生根,生根率达９４．４％。白木香试管苗移栽成活率为９３．８％。     

转基因棉花种植对根际土壤微生物群落功能多样性的影响

应用Biolog方法对两种转基因棉花及其亲本非转基因棉花根际土壤微生物的单一碳源利用水平进行了比较分析,探讨转基因棉花种植对其根际土壤微生物群落功能多样性的影响.结果表明:与非转基因亲本相比,在苗期、蕾期、吐絮期、衰老期转基因棉花种植对根际土壤微生物群落碳源利用能力、Shannon功能多样性指数和均匀度指数的影响均不显著,而在花铃期根际土壤微生物群落碳源利用能力和Shannon功能多样性指数显著降低.主成分分析表明,花铃期转基因棉花与非转基因棉花根际土壤微生物碳源利用在两主成分轴上的分异较大,碳源利用模式差异显著.     

A Comparative Test of Mechanized and Manual Transplanting of Eelgrass, Zostera marina, in Chesapeake Bay

Abstract The laborious process of manual seagrass transplanting has often limited the size of seagrass restoration efforts. This study tested the efficiency of a mechanized planting boat, previously used for transplanting Halodule wrightii, relative to manual transplanting methods for establishing Zostera marina in Chesapeake Bay. Eelgrass planting was conducted at two sites, one each in the Rappahannock and James rivers, in October 2001. The methods were evaluated by three criteria: (1) initial planting success = proportion of attempted planting units (PUs) initially established (number confirmed in sediment by divers/number attempted); (2) survival = proportion of the initially established PUs persisting over 1, 4, and 24 weeks; and (3) efficiency = labor (in person·seconds) invested in each surviving PU. Initial planting success was significantly lower for the planting boat (24 and 56% at the Rappahannock and James sites, respectively) than for manual transplanting (100% at both sites). At the Rappahannock site, survival of initially established PUs declined over time for both methods, but while mean survival was always higher for manually planted rows, differences in survival between methods were not statistically significant. At the James site, survival to 1 and 4 weeks was significantly lower for the machine than for the manual method, but survival to 24 weeks was not significantly different. While the machine was able to attempt PUs faster than the manual method (2.2 s/PU vs. 5.8 s/PU, respectively), this speed was offset by poorer planting success rates, resulting in a much greater total labor investment for each machine‐planted PU that persisted to 24 weeks than for each similarly persisting manually planted PU (40.6 person·seconds/PU and 22.4 person·seconds/PU, respectively, averaged across sites). In summary, those PUs successfully planted by the machine survived similarly to PUs planted by hand, but as a result of poorer initial planting success, the machine required a greater investment of labor and plant donor stock for each PU surviving to 24 weeks. Therefore, in its tested configuration this planting boat is not a significant improvement over the manual method for transplanting eelgrass.     

High frequency plant regeneration from protoplasts in cotton <Emphasis Type="Italic">via</Emphasis> somatic embryogenesis

A highly reproducible system for efficient plant regeneration from protoplast via somatic embryogenesis was developed in cotton (Gossypium hirsutum L.) cultivar ZDM-3. Embryogenic callus, somatic embryos and suspension culture cells were used as explants. Callus-forming frequency (82.86 %) was obtained in protoplast cultures from suspension culture cells in KM₈P medium with 0.45 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 0.93 μM kinetin (KIN), 1.5 % glucose and 1.5 % maltose. Protocolonies formed in two months with plating efficiency of 14 %. However, the callus-forming efficiencies from other two explants were low. The calli from protoplast culture were transferred to somatic embryo induction medium and 12.7 % of normal plantlets were obtained on medium contained 3 % maltose or 1 % of each sucrose + maltose + glucose, 2.46 μM indole-3-butyric acid (IBA) and 0.93 μM KIN. Over 100 plantlets were obtained from protoplasts derived from three explants. The regenerated plants were transferred to the soil and the highest survival rate (95 %) was observed in transplanting via a new method.     

资源植物罗布麻的愈伤组织诱导和植株再生研究

以罗布麻（Apocynum venetum L.）无菌苗的叶、茎和根作为外植体,在不同激素与浓度组合的MS培养基上进行愈伤组织诱导和植株再生研究。结果表明,在多个激素浓度配比的培养基中,罗布麻叶和茎的愈伤组织诱导率均可达到100%;但进一步将愈伤组织转接到含有不同激素组合的培养基进行不定芽分化时,叶和茎来源的愈伤组织的不定芽分化率有很大不同,茎来源的愈伤组织虽可在多个培养基中有不定芽分化,但最高诱导率仅达20%;而叶片来源的愈伤组织虽仅有4个激素组合培养基中有不定芽的分化,但最高分化率可达到35%。因此,构建罗布麻再生体系的最佳外植体应选择叶片。愈伤组织诱导与不定芽分化的最佳培养基均为：MS＋NAA0.1 mg/L＋6-BA 1.0 mg/L;不定芽在1/2MS＋NAA 0.2 mg/L生根培养基的生根效果最佳,不仅根群质量好,且生根率也高达100%;此再生苗的移栽成活率也最高,在适宜条件下可达75%。     

新疆棉区转Bt基因棉对棉铃虫生物学的影响

棉铃虫Helicoverpaarmigera高龄幼虫取食转Bt基因棉花组织后 ,化蛹率、羽化率、蛹重、体长均有显著下降 ,在转Bt基因棉花上棉铃虫的取食行为也受到较大的影响 ,表现为取食次数明显减少、吐丝下垂次数明显增加 ;无论是转Bt基因棉花还是常规棉花 ,棉铃虫 3龄幼虫主要分布于繁殖器官上 ,在转Bt基因棉花各繁殖器官上的分布概率为 :花 >棉铃 >棉蕾 ,常规棉花上分布概率为 :棉蕾 >棉铃 >花 ;棉铃虫高龄幼虫取食转Bt基因棉花各组织 ,成虫羽化后产卵量、卵孵化率均有明显下降 ;在较低的棉铃虫虫口密度下 ,转Bt基因棉花对棉铃虫有一定的产卵排趋性。     

石斛兰转ACS反义基因抗性原球茎及转化植株的筛选与鉴定

该试验就石斛兰转化ACS(1-氨基环丙烷-1-羧酸合成酶)反义基因的不同筛选方法和筛选处理对抗性原球茎筛选的影响,以及石斛兰转基因植株的再生与鉴定进行研究.结果表明:(1)石斛兰原球茎经带有gus报告基因和ACS反义基因的农杆菌LBA4404侵染共培养5d后除菌,采用逐渐提高选择压浓度的延迟筛选,并在低选择压浓度下切割而高选择压浓度下不切割的处理方式为抗性原球茎的最佳筛选途径,抗性原球茎获得率可达14.97％.(2)抗性原球茎繁殖时应逐渐降低选择压浓度,且在低选择压浓度下进行切割处理,繁殖倍数达到1.15倍,且原球茎生长势好.(3)抗性原球茎在1/2 MS+0.5 mg/L 6-BA培养基中的分化率达到73.85％;107株无根小苗培养于1/2 MS+1.0 mg/L NAA+50.0 mg/L Km(卡那霉素)+100.0 mg/L Cef(头孢霉素)培养基中进行生根培养,共获得了13株具有卡那霉素抗性的转化植株,转化效率达到12.15％.(4)转化植株经报告基因产物GUS组织化学检测和gus的PCR检测,证实带ACS反义基因的T-DNA已整合进石斛兰基因组中,且转基因植株在形态上与未转基因植株无明显差别,3株转基因植株移栽2个月后均已成活.     

扁桃优良砧木离体快繁

对扁桃优良砧木“Hansen”侧芽进行离体快繁研究,结果表明：外植体取材时间不同,成活率差异明显,以春季芽即将萌动的3月下旬为最佳时间,在MS 6-BA1．0mg／L GA30．2mg／L培养基上诱导芽萌发,萌发率在75％以上;以MS 6-BAl．5mg／L IAA0．1mg／L进行芽的增殖培养效果最好;1／2MS IBA0．5mg／L诱导生根,并给以黑暗预处理可使生根率达85％以上,每株生根数量多,根系质量好;采用分步炼苗移栽,提高了小苗成活率,有利于小苗后期生长。