利用植物激素调控嫁接形成的初步研究

利用黄瓜（Ｃｕｃｕｍ ｉｓｓａｔｉｖｕｓ）试管苗进行离体茎段自体嫁接,研究ＩＢＡ 和６－ＢＡ 对嫁接形成的影响时发现：进行离体茎段嫁接时,用试管苗茎段可简化嫁接过程,减少污染。嫁接茎段的颜色变化、不定根发生和愈伤组织形成与激素浓度有关。植物激素通过影响砧木和接穗间维管束桥形成的时间和数目调控嫁接组合的发育。在作者的实验中,最佳的激素条件是：在接穗培养基中加ＩＢＡ １．２ ｍ ｇ／Ｌ,在接穗和砧木培养基中加６－ＢＡ ０．３ ｍ ｇ／Ｌ。     

嫁接接合部维管组织分化的激素调节

利用黄瓜（ ＣｕｃｕｍｉｓｓａｔｉｖｕｓＬｉｎｎ．） 试管苗离体茎段自体嫁接系统, 研究ＩＡＡ和ＺＴ对砧木和接穗维管组织分化的影响, 发现外源ＩＡＡ 和ＺＴ 是砧木和接穗间维管束桥分化的必要条件。培养基中外源激素的浓度和种类通过调控维管束桥形成时间和数目以及贯通砧木和接穗的管状分子数来调节嫁接体发育。接合部维管组织分化是生长素和细胞分裂素共同作用的结果。离体茎段自体嫁接系统是一个理想的研究植物维管组织分化的新系统。     

南瓜属植物离体茎段嫁接维管组织的发育过程

在离体条件下,采用同瓜属植物的下胚轴节段进行嫁接,光镜观察发现：异种嫁接西葫芦／南瓜和南瓜自体嫁接嫁接后６－８ｄ在接穗、砧木的薄壁细胞和嫁接面处愈伤组织细胞中分化出管状分子和筛分子,边接接穗和砧木的质部和韧皮部桥在嫁接后８ｄ形成。随后随发育天数的增加其数目增加,用６（５）ＣＦ作为筛管输导的示踪剂检验了不同发育时期砧木和接穗间的连通情况,发现嫁接后８ｄ从接穗引入的６（５）ＣＦ可以输导到砧木。     

提高沙田抽茎尖嫁接成活率的研究

研究了可以影响沙田抽茎嫁接成活率的部分因素。结果表明：当沙田抽接穗选择带4个叶原基的茎尖、枳壳砧木选择黑暗培养14d的实生苗或培养基中蔗糖浓度选择7．5％时各自的嫁接成活率较高,若在茎尖嫁接切口外缠绕parafilm胶可令成活率明显增加。若在培养基中加入GA3、6－BA和IBA则明显抑制嫁接成活率。GA3、6－BA和IBA处理接穗和砧木均可提高嫁接成活率,其中以10mg/LGA3处理效果最好,嫁接成活率可达45％;6－BA和IBA处理虽然也可提高嫁接成活率,但是同时又增加了接穗的脱落死亡率。     

提高沙田柚茎尖嫁接成活率的研究

研究了可以影响沙田柚茎尖嫁接成活率的部分因素。结果表明 :当沙田柚接穗选择带 4个叶原基的茎尖、枳壳砧木选择黑暗培养 1 4d的实生苗或培养基中蔗糖浓度选择 7.5 %时各自的嫁接成活率较高。若在茎尖嫁接切口外缠绕 parafilm胶可令成活率明显增加。若在培养基中加入 GA3、6-BA和 IBA则明显抑制嫁接成活率。GA3、6-BA和 IBA处理接穗和砧木均可提高嫁接成活率 ,其中以 1 0 mg/L GA3处理效果最好 ,嫁接成活率可达 45 % ;6-BA和 IBA处理虽然也可提高嫁接成活率 ,但是同时又增加了接穗的脱落死亡率。     

西瓜嫁接体发育中木质素合成及代谢相关酶活性的变化

研究了西瓜/葫芦嫁接体发育过程中砧木和接穗部分木质素含量及其代谢相关酶的活性变化.结果表明,嫁接体发育过程中木质素生物合成加快,砧木及接穗部分的过氧化物酶(POD)、过氧化氢酶(CAT)、苯丙氨酸解胺酶(PAL)、肉桂醇脱氢酶(CAD)等的活性均明显高于对照,接穗和砧木中POD活性在15 d内持续升高,H2O2含量和CAT活性于嫁接后9~12 d出现高峰,砧木中PAL于9 d时有活性高峰而接穗一直保持较高活性而没有活性高峰,木质素含量和CAD活性持续增长,葫芦砧木木质素的代谢水平高于西瓜接穗.     

电波传递在嫁接基本理论研究中的应用

研究发现电波从接穗跃过嫁接面向砧木传递与嫁接的组织学变化相关。砧木和接穗愈伤组织细胞突破隔离层互相嵌合之前,接穗中产生的电波不能传到砧木。砧木和接穗细胞间产生次生胞间连丝后,电波即可沿接穗传至砧木,但此时传递速度慢。当砧木和接穗间产生维管束桥时,电波下传的速度加快。由此可见,通过检测电波跃过嫁接面与否及下传的速度,就可推断嫁接接合部组织学变化状况。电波传递法是一种快速鉴定嫁接植株发育过程的新方法。     

柑桔茎尖培养的初步研究

截取1～2毫米高的甜橙(Citrus sinensis)茎尖,培养子经消毒的基本培养基中。比较试验了几种植物生长调节物质、蔗糖的不同浓度和培养基的物理性状对愈伤组织发生的效应。分别观察了不同天数的离体茎尖愈伤组织发生情况。发现对愈伤组织形成较好的处理有:2,4-D0.1毫克/升+Kinetin 0.25毫克/升+NAA 2.5毫克/升,单独使用2,4-D为1.5毫克/升;蔗糖浓度为5％;以及液体培养基。将具有愈伤组织的离体茎尖,转移至激素类物质为BA 0.25毫克/升+NAA 0.1毫克/升的基本培养基中,则能促使茎芽及愈伤组织的进一步生长。但不容易形成根。然后再移入只含NAA 2毫克/升的的基本培养基中,才能分化出根,形成完整的小植株。我们已经获得暗柳、雪柑和柚子等三个柑桔品种的离体茎尖培养的植株,并揭示出在同一块离体茎尖愈伤组织生长出许多小植株的可能前景。茎尖嫁接植株是用全黑暗条件培养13～14天的枳壳(Ponciruo trifoliata)等实生苗作砧木,甜橙(Citrus sinensis)茎尖长为0.2～0.4毫米、并带叶原基2～3个作接穗,在无菌条件的实体显微解剖镜下,嫁接于砧木倒T字型的缺口上,能获得嫁接绿苗18.5％的成活率。嫁接植株培养在MS液体培养基,蔗糖浓度提高到7.5％,放置于800米烛光的荧光灯、每天光照16小时的条件下。嫁接植株生长到2～4片     

成年沙田柚树试管嫁接及微繁殖的研究

分别以两类沙田柚侧芽为接穗、酸柚为砧木进行了试管嫁接,结果表明:不同嫁接方式、接穗大小以及不同激素处理均直接影响到试管嫁接成活率.对大田侧芽而言,接穗大小以茎尖为宜,1.0mg/LGA3或0.5mg/L6-BA处理效果较佳,嫁接成活率分别达50%和30%;对侧枝沙培萌发的侧芽而言,接穗大小以茎尖 1个节间为宜,2.0mg/LGA3或0.5mg/L6-BA处理效果较佳,嫁接成活率分别达90%和80%.在相同条件下,后者的嫁接成活率一般明显高于前者.两种沙田柚嫁接苗的试管微繁殖没有明显区别,在MS 6-BA0.1mg/L培养基上均可分化不定芽,继代的嫁接成活率可达90%以上.     

砧木与接穗的相互影响

一、菊科的嫁接用刀片或锋利的刀子将向日葵(砧木)的顶部削去,然后在茎的留下部分切一纵的切口(深度4—5厘米)。接穗则用菊芋茎的上部。将它纵切成尖锐的楔形,然后作一长约5—6厘米的纵剖。半插入砧木茎的断面,一半放入带水的试管中。再用干净和浸湿的小布条将嫁接的地方包札好。摘去接穗上所有的小叶子(为了减少蒸发)。刚刚在砧本与接穗服贴后,即将试管取下,并将原在试管中的接穗茎切去。这样经过了3—4星期,接穗与砧木结合得很好的时候。即     

Successful nodulation of Casuarina by Frankia in axenic conditions

AIMS: In order to depict the fine interactions that lead to nodulation, absolute microbiological control of the symbiotic partners is required, i.e. the ability to obtain in vitro axenic nodulation, a condition that has never been fulfilled with the Casuarina-Frankia symbiosis. The effects of culture conditions on plant growth and nodule formation by Casuarina cunninghamiana were investigated. METHODS AND RESULTS: Axenic (capped tubes with different substrates), and nonaxenic cultures (Gibson tubes, pot cultures) were tested. In axenic conditions, C. cunninghamiana, inoculated with Frankia, had poor growth and did not form nodules at 6 weeks. Plants cultivated in Gibson tubes reached the four axillary shoots stage within 6 weeks and formed nodules 4 weeks after inoculation. Sand-pot cultures allowed us to relate the plant development stage at inoculation with nodulation. CONCLUSIONS: The sterile replacement of the cap by a plastic bag increased plant growth and enabled nodule formation 6 weeks after inoculation. The new system of plant culture allows the axenic nodule formation 6 weeks after inoculation. Nodulation behaviour is related to plant development and confinement. SIGNIFICANCE AND IMPACT OF THE STUDY: This axenic plant nodulation system is of major interest in analysing the roles of Frankia genes in nodulation pathways.     

Programmed cell death and tissue remodelling in plants

The use of programmed cell death (PCD) to remodel plants at the cellular, tissue, and organ levels is particularly fascinating and occurs in such processes as tracheary element differentiation, lysigenous aerenchyma formation, development of functionally unisexual flowers from bisexual floral primordia, and leaf morphogenesis. The formation of complex leaf shape through the use of PCD is a rare event across vascular plants and occurs only in a few species of Monstera and related genera, and in the lace plant (Aponogeton madagascariensis). During early development, the lace plant leaf forms a pattern of equidistantly positioned perforations across the surface of the leaf, giving it a lattice-like appearance. Due to the accessibility and predictability of this process, the lace plant provides highly suitable material for the study of developmentally regulated PCD in plants. A sterile lace plant culture system has been successfully established, providing material free of micro-organisms for experimental study. The potential role of ethylene and caspase-like activity in developmentally regulated PCD in the lace plant is currently under investigation, with preliminary results indicating that both may play a role in the cell death pathway.     

Phytoremediation of ethion by water hyacinth (Eichhornia crassipes) from water

The potential of water hyacinth (Eichhornia crassipes) to remove a phosphorus pesticide ethion were investigated. The disappearance rate constants of ethion in culture solutions were 0.01059, 0.00930, 0.00294, and 0.00201 h-1 for the non-sterile planted, sterile planted, non-sterile unplanted, and sterile unplanted treatment, respectively, which were significantly different and implied that plant uptake and phytodegradation contributed 69% and that of microbial degradation took up 12% to the removal of the applied ethion. The accumulated ethion in live water hyacinth plant decreased by 55-91% in shoots and 74-81% in roots after the plant growing 1 week in ethion free culture solutions, suggesting that plant uptake and phytodegradation might be the dominant process for ethion removal by the plant. This plant might be utilized as an efficient, economical and ecological alternative to accelerate the removal and degradation of agro-industrial wastewater polluted with ethion.     

Competition ofRhizobium strains in nodule-formation

Summary By the use of a sterile culture system experiments have been carried out with peas in order to elucidate to what extent inoculation with an ineffective bacterial strain effects the ability of an effective strain to produce further nodulation. It has been established that effective strains, when applied to the plant after the formation of the first nodules by an ineffective strain, altogether fail to form nodules or the nodulation is delayed very much. Differences are noted in this respect between different bacterial strains. The results can be explained easiest by assuming that the strain first entering the roots causes an immunity in the plant against later infections. Other possibilities for explanation are also discussed. The conception that the resistance would be due to the saturation of roots with nodules formed by the first strain which would inhibit further nodulation is not in accordance with the results recorded.A method has been introduced for cultivation of plants under sterile conditions by dividing the roots into two or more culture flasks. This method is of great value when the nutrient uptake of plants is elucidated. Successive inoculation with ineffective and effective strains gave varying results by this technique. The experiments seem to imply that if only a part of the root system is inoculated with an ineffective strain, immunity does not regularly occur against later infections by effective strains. This suggests that the immunity is a somewhat local phenomenon.     

不同培养方法对甘薯无菌苗生长的影响

试验结果表明,在甘薯组织培养中液体培养不降低无菌苗生长势,对无菌苗生长有一定促进作用,适合于甘薯无菌苗的快速繁殖。液体培养20d比相同条件下固体培养的无菌苗植株高度高出63．16％,经方差分析,差异显著,具有统计学意义;液体培养60d比相同条件下固体培养的无菌苗植株高度,单株叶片数,单叶面积分别高出59．43％,106．70％,49．82％,经方差分析,前两者差异显著,具有统计学意义。     

Long-term culturing of plants with aseptic roots Determination of rape root exudates

Abstract A plant culture system allowing growth of aseptic roots has been designed. One version of the system comprises vessels developed for plant root-microorganism interaction studies in series. A second version has been built for measurements of different physiological parameters of the shoot and root system during growth periods of at least 2 months. The system was tested by determining soluble organic carbon glucose and sucrose in the root exudates of rape plants (Brassica napus L.) grown for 3 weeks in the culture system. The plants were cultivated with a sterile or a non-sterile root system, or with a root system infected with Verticillium dahliae Kleb.     

Establishment, graft union characteristics and growth of Prunus micrografts

A micrografting technique for use on shoots derived by shoot-tip culture is described. Autografts of Prunus domestica cv. Hauszwetsche as well as heterografts of several sour cherry cultivars ( Prunus cerasus L. cvs Schattenmorelle, Weiroot 158, Köröser) were established. Successful graft formation in vitro was confirmed by translation of ⁸⁶Rb⁺ from the stock root into growing scion tissues. A mechanically strong graft union was formed during the course of a 3-week subculture of micrografts in a liquid medium without the addition of growth regulators. In the case of graft rejection, ⁸⁶Rb⁺ was mainly attracted to new developing shoots from lateral meristems of the stock plant. Histological examination of the graft union revealed callus formation, cytodifferentiation and xylogenesis leading to the formation of vascular connections. Stem elongation after micrografting was related to vigour of the stock and scion genotypes. Early stem elongation could be used as a criterion for preselection of growth vigour in graft combinations. Micrografts were transferred to soil and grown in the greenhouse.     

The growth and phenotypic expression of human osteoblasts

The care of patients with a skeletal deficiency currently involves the use of bone graft or a non-biologic material such as a metal or polymer. There are alternate possibilities in development which involve the growth of bone cells (osteoblasts) on degradable polymer scaffolds. These tissue engineering strategies require production of the polymeric scaffold, cellular harvest followed by either ex vivo or in vivo growth of the cells on the scaffold, and exploration of the interaction between the cell and scaffold. Research into these strategies utilizes cells from a variety of species, but clinical applications will likely require human osteoblasts. This study explores the process whereby human osteoblasts are harvested under sterile conditions during joint replacement surgery from normally discarded cancellous bone, transported from the operating room to the lab, and grown in culture. This process is feasible, and the cells express their phenotype via the production of alkaline phosphatase and collagen in culture.     

Development of an automated plant culture system

An apparatus was constructed that could be used to grow plant tissues, organs, and whole plantlets under sterile conditions. This system accommodated independent or multiple concurrent growth of cultures. Growth of plants either equalled or exceeded that observed using the manual transfer procedure. The automated plant culture system (APCS) consists of silicone tubing, 2 impeller pumps, 2 glass medium reservoir bottles, a 3-way stainless steel valve, a plant culture chamber, and an interface module containing relay boards. Control of the APCS is through interfacing with a microcomputer (e.g. Apple IIe or Atari 400). The computer controlled medium introduction, evacuation, and replenishment in a sterile environment. The APCS was inexpensively constructed and provides a labor-saving, long-term method to culture plants in vitro.Mention of a trade name or proprietary product in the paper does not constitute a guarantee or warranty of the product by the US Department of Agriculture and does not imply its approval to the exclusion of other products that may also be suitable.     

鸟巢蕨孢子繁殖技术研究

研究了鸟巢蕨孢子无菌播种和常规播种两种繁殖方法。结果表明,鸟巢蕨无菌播种中孢子萌发率最高可达66.7%,原叶体在固体培养基上培养不能诱导出孢子体,而需经过振荡培养后才能诱导出孢子体;常规播种法更容易诱导出孢子体,每盆播0.02 g孢子时,每克孢子可产生孢子体4 000株以上,比无菌播种操作简便,成本低。因此,孢子常规播种法更适合于鸟巢蕨规模化生产。