Induction of tumours by various strains of Agrobacterium tumefaciens on Abies nordmanniana and Picea abies

Abies nordmanniana and Picea abies seedlings were inoculated with a wild‐type (C 58) or an attenuated strain (rooter or shooter mutants) of Agrobacterium tumefaciens. Large tumours were formed on A. nordmanniana in response to the wild‐type and rooter strains, 60–65 % of the seedlings being susceptible. Smaller tumours were formed on Picea abies in response to the wild‐type strain, at least 12% of the seedlings being susceptible. Tumours from both species induced by the wild‐type strain grew in vitro on medium without added phytohormones.     

农杆菌介导BADH基因转化美丽胡枝子的研究

以子叶节为外植体,草丁膦为选择剂,对根癌农杆菌LBA4404介导的美丽胡枝子遗传转化进行研究,建立美丽胡枝子遗传转化体系,获得转BADH基因植株,最高转化率可达28%.抗性植株经分子检测表明,目的基因已整合进美丽胡枝子基因组中.NaCI胁迫下,转基因植株能积累更多的甜菜碱,证明BADH基因在转化植株中得到正常表达.     

Regeneration and gene transformation systems of Robinia pseudoacacia ''Idaho'' mediated by Agrobacterium tumefaciens

Robinia pseudoacacia 'Idaho' is one of several multi-purpose trees used in ornamental, soil and water conservation, fodder and nectar sources. Plant abiotic stress tolerance transformed by genes could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration. For this paper, we studied the effects of auxin and cytokine on Idaho locust in vitro regeneration and the establishment of gene transformation systems for plants mediated by Agrobacterium tumefaciens. Results showed that the ratios of cytokinin and auxin were the major factors affecting adventitious bud differentiation on a MS medium; the concentration of 0.5inhibit rooting. The most effective antitoxin for screening transgenic Idaho locust shoots was G418 and the most sensitive concentration of it was 8 mg·L-1.     

Regeneration and gene transformation systems of Robima pseudoacacia ''''Idaho'''' mediated by Agrobacterium tumefaciens

Robinia pseudoacacia 'Idaho' is one of several multi-purpose trees used in ornamental, soil and water conservation, fodder and nectar sources. Plant abiotic stress tolerance transformed by genes could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration. For this paper, we studied the effects of auxin and cytokine on Idaho locust in vitro regeneration and the establishment of gene transformation systems for plants mediated by Agrobacterium tumefaciens. Results showed that the ratios of cytokinin and auxin were the major factors affecting adventitious bud differentiation on a MS medium; the concentration of 0.5 mg·L-1 6-BA benefitted callus proliferation and 0.25 mg·L-1 IBA promoted shoot rooting; however, a higher IBA concentration will inhibit rooting. The most effective antitoxin for screening transgenic Idaho locust shoots was G418 and the most sensitive concentration of it was 8 mg·L-1.     

Genetic transformation of loblolly pine using mature zygotic embryo explants by Agrobacterium tumefaciens

Introduction1Genetictransformationinconifershasthepotentialtoallowtheselectiveimprovementofindividualtraitsinelitecloneswhilestillmaintainingtheexistingcombinationofgenesresponsibleforthesuperiorphenotype(Charestetal.1991;Jamesetal.1996;Walteretal.1999).Atpresent,althoughconsiderableresearchefforthasbeendevotedtothegeneticengineeringofconiferspecies(Sederoffetal.1986;Bekkauoietal.1988,1990;Robertsonetal.1992;Bomminenietal.1993;Shinetal.1994;Klimaszewskaetal.1997),ithaslaggedbehindadvancesma…     

根癌农杆菌介导Caz基因转化烟草条件研究

利用根癌农杆菌介导Caz基因转化烟草,将外源基因导入烟草愈伤组织,建成可诱导表达细胞质和细胞核CaM/CaM结合多肽的转基因烟草．研究结果表明:植物材料的选择和培养、农杆菌的培养和稀释、植物材料与农杆菌共培养的程度及转基因植株的筛选条件是影响农杆菌转化效率的重要因素．     

Lightwood induction in Pinus sylvestris by means of mechanical wounding

In a field experiment the influence of different girdling patterns like wide girdling, longitudinal debarking, debarking with one or several bridges, narrow girdling and bark crushing on resin soaked wood formation (lightwood) in Pinus sylvestris L. trees was investigated. Four years after treatment the concentrations of total extractives and resin acids were measured. It is shown that treatment methods differently affected concentrations of total extractives and resin acids. Longitudinal debarking with a wide bridge and debarking with several narrow bridges were the most successful methods for lightwood induction.     

Regeneration and gene transformation systems of Robinia pseudoacacia ‘Idaho’ mediated by Agrobacterium tumefaciens

Robinia pseudoacacia ‘Idaho’ is one of several multi-purpose trees used in ornamental, soil and water conservation, fodder and nectar sources. Plant abiotic stress tolerance transformed by genes could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration. For this paper, we studied the effects of auxin and cytokine on Idaho locust in vitro regeneration and the establishment of gene transformation systems for plants mediated by Agrobacterium tumefaciens. Results showed that the ratios of cytokinin and auxin were the major factors affecting adventitious bud differentiation on a MS medium; the concentration of 0.5 mg•L^–1 6-BA benefitted callus proliferation and 0.25 mg•L^–1 IBA promoted shoot rooting; however, a higher IBA concentration will inhibit rooting. The most effective antitoxin for screening transgenic Idaho locust shoots was G418 and the most sensitive concentra-tion of it was 8 mg•L^–1.     

不同因素对农杆菌介导的杉木转化效率的影响

以杉木试管苗靠近茎尖的茎段为受体材料,研究6个影响杉木遗传转化效果的因素,初步确定杉木茎段较为合适的转化体系为:预培养1～3 d,OD600 nm为0.1～0.4的菌液浸染10～15 min,共培养3～5 d,共培养基附加乙酰丁香酮(AS)80 μmol·L-1,共培养后延迟3 d筛选.在初次筛选培养基上共得到186个卡那霉素(Km)抗性芽,在二次筛选培养基上获得39个Km抗性芽.PCR检测有2个Km抗性芽呈稳定阳性,初步证明外源天麻抗真菌蛋白(GAFP)基因已整合到这2个Km抗性芽的基因组DNA中.     

根癌农杆菌对感染植原体的泡桐组培苗症状的影响

采用含有激素合成相关基因的根癌农杆菌，伤口接种已感染植原体的泡桐丛植组培苗和健康组培苗，结果发现对丛植苗的致瘤能力明显低于健康对照苗，且被接种病苗的丛枝症状缓解，从健苗获得的T-DNA转化泡桐瘤组织细胞能在无激素培养基上稳定生长和连续继代培养2年以上，说明瘤组织细胞自身已获得了细胞分裂素和生长素合成能力，根据已报道的根癌农杆菌株系pTil5955T-DNA的异戊烯基转移酶基因（ipt)的保守序列，设计了一对引物（CYT和CYT′），用多聚酶链式反应（PCR）扩增了我国杨树致瘤农杆菌ipt基因部分序列（427bp片段），也从遗传转化的两个泡桐无性系瘤组织At-ZH和At-T35扩增出此特异片估，从而进一步肯定了T-DNA已被整合到泡桐的染色体上表明泡桐易于通过Ti质粒载体途径进行基因转移操作，但用此引物未能从泡桐、甘薯健株和感染植原体的组培病苗扩增出相应的427bp特异片段，当用此遗传转化瘤组织嫁接病苗时，可减轻从枝症状的严重度，延长病苗的存活时间和诱导病株生根，这进一步证实了泡桐在与植原体相互作用过程中激素代谢发生了变化。     

抗生素对农杆菌的抑制和杨树叶片分化的影响

中嘉8号杨为湖北省林业厅和中国林科院从美洲黑杨的实生后代中选育的杨树新品系。以中嘉8号杨为材料,探讨了几种抗生素对农杆菌的抑制和对杨树离体叶片分化的影响。结果表明,噻孢霉素的抑菌效果最好,选用100~200 mg/L的噻孢霉素来抑菌是比较合适的,羧苄青霉素和链霉素对杨树叶片离体培养的毒性比噻孢霉素大,卡那霉素完全抑制芽的分化。     

人工诱导阔叶红松林改良土壤效果分析

文章通过比较不同类型红松针阔混交林和红松纯林的林分枯落物含量、土壤理化性质及土壤酶活性等性状可知,单位面积枯落物总量红松针阔混交林比红松纯林高40%~60%,其半分解的枯落物量均高于未分解的枯落物量,而红松纯林则相反;红松针阔混交林中枯落物的持水量较红松纯林高26%~75%,养分含量明显高于红松纯林;红松针阔混交林土壤理化性质及酶活性都明显优于红松纯林。     

Regeneration of phenotypically normal English elm (Ulmus procera) plantlets following transformation with an Agrobacterium tumefaciens binary vector

A transformation system was developed for English elm (Ulmus procera Salisbury) using Agrobacterium tumefaciens C58 pMP90 p35SGUS/INTRON, allowing for the transfer of foreign genes and regeneration of phenotypically normal elm plantlets. The PCR analysis indicated that both nptII and uidA genes were stably inserted in the plant genome. beta-Glucuronidase histochemical and fluorimetric assays revealed expression of the uidA gene in the shoots, leaves, stems and roots of regenerated transgenic plants. The DNA-DNA hybridizations confirmed the presence of the uidA gene in regenerant plants. Factors influencing successful transformation and regeneration of elms included: identifying gene-transfer-proficient Agrobacterium strains for use with elms; developing an infection protocol allowing T-DNA transfer while retaining the ability to remove inciting bacteria; and identifying selection conditions to eliminate non-transformed material and choice of regeneration medium to allow shoot production. The potential utility of an effective elm transformation and regeneration system in the control of Dutch elm disease is discussed.     

美国黄松离体胚培养条件下不定芽的形成与根产生的研究

以美国黄松成熟胚为外植体在MS、GD、SH和N6 培养基上诱导不定芽 ,试验结果表明 ,基本培养基的种类对外植体不定芽的诱导起主要作用 ,GD最好 ,SH次之 ,MS和N6 最差。GD 0 5mg·L- 1 6 BA ,对外植体不定芽的诱导率达 5 5 % ,平均增殖率为 6 ,最大增殖率达 10 ;NAA不利于外植体不定芽的诱导 ;培养基中加入适量的活性炭有利于不定芽的形成和生长。对不定芽在GD、SH和 1 2SH培养基上进行生根诱导 ,试验结果表明 ,基本培养基的种类对不定芽形成根起主要作用 ,GD、SH不能诱导不定芽生根 ,1 2SH可以使不定芽生根 ,其对不定芽的诱导率为 2 2 % ,1 2SH NAA 0 5mg·L- 1 对不定芽生根的诱导率为 3 3% ;NAA对不定芽生根具有促进作用。在离体培养条件下 ,以美国黄松种胚为外植体获得了再生小植株     

马尾松苦竹混交林根系分布格局

对马尾松苦竹混交林根系分布格局进行研究.结果表明:混交林根系垂直分布比较合理.水平上有重叠,但根系多相互交错,穿插延伸.低密度混交经营的马尾松细根在40～60 cm土层占细根总量的83.8%,而苦竹竹鞭及其构成的竹林地下吸收、输导、贮存系统主要分布在0～40 cm土层占96.0%,较合理地利用了不同土层的营养物质.混交林分中马尾松水平根幅8.9 m,但在0～40 cm土层主要为水平骨骼根,呈疏散框架扩展延伸,给苦竹鞭根的运行、穿插腾出了空间.而苦竹的竹根水平占据空间较小,一般在30 cm左右,鞭根稀少.马尾松利用疏林结构模式兼营苦竹的混交林分不仅形成合理的地上结构,而且地下结构也较合理.     

马尾松人工林根呼吸的季节变化及影响因子

2007年1月至2008年12月,以长沙天际岭国家森林公园马尾松群落为研究对象,采用挖壕法研究马尾松群落去除根系后土壤呼吸速率动态及其与土壤温、湿度的相关关系。结果表明:马尾松群落和去除根系处理的土壤呼吸年变化范围分别为0.29～3.19,0.25～2.33μmol·m-2s-1,年均分别为1.56,1.03μmol·m-2s-1。去除根系土壤呼吸速率比对照降低12.2%～55.1%,根呼吸年均贡献率为28.3%。马尾松群落和去除根系的土壤呼吸速率与5cm土壤温度之间均呈显著指数相关,温度敏感系数Q10值分别为2.10和1.82,估算出根呼吸的Q10值为2.94。马尾松群落和去除根系的土壤呼吸速率与土壤湿度之间相关关系均不显著(P﹥0.05),但根呼吸与土壤湿度之间呈显著线性相关(P=0.023)。     

Selection of culture conditions for callus induction and proliferation by somatic embryogenesis of Pinus koraiensis

The induction and proliferation of embryogenic callus are key steps for large-scale propagation of somatic embryogenesis pathway and long-term preservation of coniferous germplasm.Callus can be induced from immature embryos of Korean pine(Pinus koraiensis Sieb.et Zucc.;Pinaceae) as explants,but there are problems,such as low proliferation efficiency,loss of embryogenicity,poor vigor;thus,best conditions for proliferation and culture of immature embryos of Korean pine are not yet clear.To solve the problems with somatic embryogenesis of Korean pine and determine the best culture conditions for callus induction and proliferation,we varied hormone concentration,subculture cycle of proliferation and other plant growth regulators combinations in media to induce callus formation by megagametophytes of three Korean pine families at different developmental stages,then analyzed the effects on embryogenic callus retention and cell proliferation using a quadratic regression orthogonal rotation design.The results showed that the family origin and collection date of explants significantly affected callus induction(induction rate reached1.67%).Embryogenic maintenance and callus proliferation were best on DCR medium supplemented with 0.25 mg L~(-1)6-benzyl adenine,1 mg L~(-1) naphthaleneacetic acid,30 g L~(-1)sucrose,500 mg L~(-1),L-glutamine,500 mg L~(-1) casein hydrolysis and 6.5 g L~(-1) agar.In addition,the combination of 2,4-dichlorophenoxyacetic acid+6-benzyl adenine also had a better proliferative effect on callus.The effects of different combinations of growth regulators on callus proliferation efficiency were significantly different.Transfer to new medium every 13-15 days not only maintained robust callus vigor,but also yielded a larger proliferation coefficient.The techniques and conditions for embryogenic callus induction and proliferation of Korean determined here will serve as a foundation for establishing a large-scale system for somatic embryogenesis and propagation of Korean pine.     

不同光照条件下红松幼苗光合作用诱导反应的研究

无论光通量密度(PPFD)为200靘ol穖-2穝-1还是500靘ol穖-2穝-1,生长在林冠下的红松幼苗光合速率达稳定净光合速率的50%和90%的诱导时间均长于全光条件下生长苗木。全光下生长的红松幼苗在PPFD为500靘ol穖-2穝-1时的诱导反应较PPFD为200靘ol穖-2穝-1时缓慢,而林冠下生长苗木则恰好相反。表明林冠下生长红松幼苗的光合作用诱导期与光斑强度有关。图3参6。