Agrobacterium tumefaciens-mediated GUS gene transformation of Robinia pseudoacacia ‘Idaho’

Based on the plant regeneration system, a GUS gene transformation system to Idaho locust (Robinia pseudoacacia ‘Idaho’) mediated by Agrobacterium tumefaciens was established. The successful transformation was confirmed by regenerating the shoots from the infected leaves in the presence of hygromysin; by histochemical X-gluc assays of β-glucuronidase (GUS) and by PCR and PCR-Southern blotting analysis. The ratio of positive transgenic plants is 5.8% (5 out of 86 plants). With this system, the target gene DREB was introduced into the leaves of Idaho locust. The transgenic plants regenerated, which was verified by PCR-Southern blot-ting. It is suggested that the transformation system could be a new, simple, reliable and practical route to gene transformation of R. pseudoacacia ‘Idaho’ mediated with A. tumefaciens.     

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.     

Agrobacterium tumefaciens-mediated GUS gene transformation of Robinia pseudoacacia ''''Idaho''''

Based on the plant regeneration system, a GUS gene transformation system to Idaho locust (Robinia pseudoacacia 'Idaho') mediated by Agrobacterium tumefaciens was established. The successful transformation was confirmed by regenerating the shoots from the infected leaves in the presence of hygromysin; by histochemical X-gluc assays ofβ-glucuronidase (GUS) and by PCR and PCR-Southern blotting analysis. The ratio of positive transgenic plants is 5.8% (5 out of 86 plants). With this system, the target gene DREB was introduced into the leaves of Idaho locust. The transgenic plants regenerated, which was verified by PCR-Southern blotting. It is suggested that the transformation system could be a new, simple, reliable and practical route to gene transformation of R. pseudoacacia 'Idaho' mediated with A. tumefaciens.     

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.     

Determination of drought tolerance using root activities in Robinia pseudoacacia ‘Idaho’ transformed with mtl-D gene

Idaho locust (Robinia pseudoacacia ‘Idaho’) is an exotic multi-purpose tree used in landscaping, soil and water conservation, fodder sources and others. To improve its drought tolerance for reclaiming arid land, five lines of transformed mtl-D gene, as osmotic regulator in plant cells, have been selected and managed to determine their drought tolerance under experimental conditions. Qualitative and quantitative variables of transformed plants were studied. The critical value of drought tolerance was determined by detecting the 2,3,5-triphenyl tetrazolium chloride (TTC) reductants in roots and soil water content (SWC). The critical value for drought tolerance was SWC 6% while for the control plants the critical SWC was 8%; a moderate level of SWC is 13% and the highest SWC for plant endurance was 18%. The method proved to be reliable and sensitive in the evaluation of drought tolerance for forest trees.     

Rooting and acclimatization of in vitro plantlets of mtl-D gene modified Robinia pseudoacacia ‘Idaho’

Robinia pseudoacacia 'Idaho' (Robinia×ambigua 'Idahoensis', R. pseudoacacia×R. viscosa) modified by a mtl-D gene went through five lines and had characteristics of drought tolerance. Three stages of their micropropagation had been studied by previous investigators. The other two stages, in vitro shoot rooting and plantlet acclimatization, still remained unsolved in the laboratory. For this paper, we studied the later two stages based on the previous achievements. Results showed that the highest rooting rate of Idaho locust was 98.4% when the in vitro shoots, over 2.5 cm in height and 0.08 cm in diameter, were placed on a half strength MS basal medium with 0.4 mg·L-1 IB A and 0.1 mg·L-1 NAA as supplements and were solidified with 0.5% agar; the highest survival rate was 98.3% when the rooted plantlets were potted in vermiculite. All the stages for micropropagation of the Idaho locust, modified by the mtl-D gene, were assembled completely. The tissue culture plants grow well in the field.     

Determination of drought tolerance using root activities in Robinia pseudoacada ''''Idaho'''' transformed with mtl-D gene

Idaho locust (Robinia pseudoacacia 'Idaho') is an exotic multi-purpose tree used in landscaping, soil and water conservation, fodder sources and others. To improve its drought tolerance for reclaiming arid land, five lines of transformed mtl-D gene, as osmotic regulator in plant cells, have been selected and managed to determine their drought tolerance under experimental conditions. Qualitative and quantitative variables of transformed plants were studied. The critical value of drought tolerance was determined by detecting the 2,3,5-triphenyl tetrazolium chloride (TTC) reductants in roots and soil water content (SWC). The critical value for drought tolerance was SWC 6% while for the control plants the critical SWC was 8%; a moderate level of SWC is 13% and the highest SWC for plant endurance was 18%. The method proved to be reliable and sensitive in the evaluation of drought tolerance for forest trees.     

Determination of drought tolerance using root activities in Robinia pseudoacacia ''Idaho'' transformed with mtl-D gene

Idaho locust (Robinia pseudoacacia 'Idaho') is an exotic multi-purpose tree used in landscaping, soil and water conservation, fodder sources and others. To improve its drought tolerance for reclaiming arid land, five lines of transformed mtl-D gene, as osmotic regulator in plant cells, have been selected and managed to determine their drought tolerance under experimental conditions.Qualitative and quantitative variables of transformed plants were studied. The critical value of drought tolerance was determined by detecting the 2,3,5-triphenyl tetrazolium chloride (TTC) reductants in roots and soil water content (SWC). The critical value for drought tolerance was SWC 6% while for the control plants the critical SWC was 8%; a moderate level of SWC is 13% and the highest SWC for plant endurance was 18%. The method proved to be reliable and sensitive in the evaluation of drought tolerance for forest trees.     

Establishment of a transgenic system in fast-growing black locust (Robinia pseudoacacia L.)

The AhDREB1 gene, cloned from Atriplex hortensis L., was transferred into black locust (Robinia pseudoacacia L.) by an Agrobacterium-mediated transformation. The results suggest that stems of black locust sub-cultured in vitro for 20 d are suitable for genetic transformation. The optimum concentrations of kanamycin and cefotaxime were 30 and 150 mg.L-1, respectively. Impor-tant factors affecting the transformation efficiency were studied by means of a L9(34) orthogonal design. An effective system for ge-netic transformation in black locust was developed as follows: the stems were pre-cultured for 2 d, immersed in the Agrobacterium solution (OD6oo = 0.7) with 10 mg'L-1 acetosyringone for 21 min and then co-cultured for 2 d. The selection pressures, changing from low to high, could improve transformation efficiency. The transgenic plants were identified by a PCR method. The PCR results indicated that the AhDREB1 gene had been integrated into the genome of black locust and two lines of the transgenic plants were obtained.     

Establishment of a transgenic system in fast-growing black locust （Robinia pseudoacacia L.）

The AhDREB1 gene, cloned from Atriplex hortensis L., was transferred into black locust （Robiniapseudoacacia L.） by an Agrobacterium-mediated transformation. The results suggest that stems of black locust sub-cultured in vitro for 20 d are suitable for genetic transformation. The optimum concentrations of kanamycin and cefotaxime were 30 and 150 mg.L-1, respectively. Important factors affecting the transformation efficiency were studied by means of a L9（3^4） orthogonal design. An effective system for genetic transformation in black locust was developed as follows： the stems were pre-cultured for 2 d, immersed in the Agrobacterium solution （OD600 = 0.7） with 10 mg·L^-1 acetosyringone for 21 min and then co-cultured for 2 d. The selection pressures, changing from low to high, could improve transformation efficiency. The transgenic plants were identified by a PCR method. The PCR results indicated that the AhDREB1 gene had been integrated into the genome of black locust and two lines of the transgenic plants were obtained.     

谷胱甘肽合成酶基因StGCS-GS的表达提高大肠杆菌对重金属的抗性

本文将来源于嗜热链球菌（S.thermophilus）的谷胱甘肽双功能合成酶基因（StGCS-GS）通过载体pETG-A10转入大肠杆菌中。在IPTG诱导后,表达产物通过凝胶电泳（SDS-PAGE）检测,重组后的融合蛋白His-GCS-GS用His-镍蛋白纯化柱纯化,SDS-PAGE和western blot检测纯化效果,鉴定表达产物。检测结果与预期一致。为了研究StGCS-GS基因的表达对大肠杆菌在重金属胁迫下的抗性作用,实验对转化大肠杆菌进行不同浓度Cd2＋、Ni2＋、Cu2＋胁迫的耐受性测试。结果发现,在1mM CdCl2、2mM NiSO4胁迫条件下,转化大肠杆菌的生长状态明显优于对照,但在Cu2＋胁迫条件下,两者没有明显区别。表明StGCS-GS基因的表达能够提高大肠杆菌对Cd2＋、Ni2＋的抗性,在其逆境胁迫下的生长发挥重要作用。     

湖南桃江黄脊竹蝗产卵量观察与研究

通过抽样调查黄脊竹蝗卵块卵粒数、解剖雌成蝗每次怀卵卵粒数、笼养观察成蝗产卵次数与每次产卵量，经数据分析估测桃江县自然界竹蝗产卵量为22～132粒、1～6次（块）。一般年份产卵量为44～66粒、2—3次。     

抗虫转基因杨树的培育及其抗虫性初步研究

以杨树优良新品种欧美杨107(Populus×euramericanacl."74/76")为试材,通过根癌农杆菌(Agrobacteriumtumefaciens)介导法,将Bt毒蛋白基因导入107杨的叶片及茎段外植体,经潮霉素抗性筛选,获得了转基因再生植株。提取转基因植株的总DNA,经PCR扩增,部分植株呈阳性反应,证明目的基因已经整合到杨树基因组中。以转基因杨树叶片饲喂天幕毛虫幼虫的杀虫试验结果表明,转基因杨树表现出一定的杀虫活性。     

竹蝗的生物防治研究

采用类产碱假单胞菌（ Ｐｓｅｕｄｏｍ ｏｎａｓ ｐ ｓｅｕｄｏａｌｃａｌｉｎｇｅｎｃｅｓ）,与苏菌（ Ｂａｃｉｌｌｕｓ ｔｈｕｒｉｎｇｉｅｎｓｉｓ）组合制成生物灭蝗剂,在竹蝗防治上取得了较好的防治效果。几年来,防治面积近 １ ０００ｈｍ ２,防效（致死率）在 ８０％ 左右,减少了竹业的经济损失,并维护了林区的生态平衡。     

ACC氧化酶cDNA克隆及其对美洲黑杨体内乙烯产生的反义抑制

利用ＲＴ－ＰＣＲ技术克隆了番茄ＡＣＣ氧化酶基因ＬＥＥＴＨＹＢＲ编码区０．９ｋｂ的ｃＤＮＡ片段,经酶切图谱分析和序列分析鉴定后,反向插入到植物表达载体ｐＢｉｎ４３８中,构建了表达ＡＣＣ氧化酶反义ＲＮＡ的二元载体。用农杆菌侵染美洲黑杨叶片,在含卡那霉素的ＭＳ培养基上选择转化子和植株再生,通过ＰＣＲ检测筛选到１６株转基因杨树植株,Ｓｏｕｔｈｅｒｎｂｌｏｔ分析初步确证了外源基因是以单拷贝插入到杨树基因组中;对杨树幼苗乙烯释放量的测定结果表明转基因杨树的乙烯释放量为对照植株的２８％。     

球毛壳菌(Chaetomium globosum)小热休克蛋白HSP22.4基因克隆及表达

小热休克蛋白(sHsps_)是生物体中参与防御反应的内源细胞保护因子。为了研究球毛壳菌中(Chaetomiumglobosum)中的小热休克蛋白(AY491980)。对球毛壳菌的小热休克蛋白进行了克隆和在大肠杆菌中进行了表达。BlastX分析表明球色壳菌的小热休克蛋白与粗糙脉孢菌(Neurosporacrassa)小热休克蛋白在氨基酸上的序列同源性最高,为65%。将HSP22.4基因编码区插入原核表达载体pGEX-4T-2、构建成表达质粒pGEX-HSP。利用pGEX-HSP质粒转化大肠杆菌BL21,IPTG诱导后SDS-PAGE检测表达情况,结果发现在50kD处有一特异性融合蛋白带,与预期相符,说明HSP22.4基因已经在大肠杆菌中表达。本研究为进一步研究小热休克蛋白的功能奠定了基础。图6参15。     

Black locust (Robinia pseudoacada L.) clonal seed orchards in Hungary

Black locust (Robinia pseudoacacia L.) is one of the most important stand-forming tree species in Hungary and its importance is increasing in many countries. The main aim of the discussed new selection programme is to identify black locust clones with good performance and good form for setting up clonal seed orchards. As a result of selection programme 16 new black locust clones have been improved. In spring 2002 a black locust seed orchard was established with the newly selected clones. About 40% of the plants can be considered to belong to the height growth rate class 1 and 2. Hungary was the first country where micropropagated black locust planting material was used for seed orchard establishment.     

Black locust (Robinia pseudoacacia L.) clonal seed orchards in Hungary

Black locust (Robinia pseudoacacia L.) is one of the most important stand-forming tree species in Hungary and its importance is increasing in many countries. The main aim of the discussed new selection programme is to identify black locust clones with good performance and good form for setting up clonal seed orchards. As a result of selection programme 16 new black locust clones have been improved. In spring 2002 a black locust seed orchard was established with the newly selected clones. About 40% of the plants can be considered to belong to the height growth rate class 1 and 2. Hungary was the first country where micropropagated black locust planting material was used for seed orchard establishment.     

Assessing Forest Health Conditions in Idaho with Forest Inventory Data

Periodic inventory measurements of forest characteristics published by the U.S. Forest Service are used to assess trends in forest conditions in Idaho. Forest species composition, measured by growing stock volume, has changed since 1952. Western white pine and ponderosa pine have declined by 60% and 40%, respectively. True firs (mostly grand fir) increased by 60% lodgepole pine by almost 40%, and Douglas-fir, the predominant species throughout the state, increased by 15%. Measurements of tree mortality across the state and region from 1952 to 1987 establish a baseline regional range for judging current conditions. Recent mortality data from some Idaho national forests are much higher than the upper limit of the baseline regional range. On the Boise and Payette National Forests in southwestern Idaho, annual mortality exceeds annual growth. Recent inventories of national forests in northern Idaho show mature stands have mortality well above the baseline regional range, which projects into a negative net growth situation. Recent inventories of private and other public forests in northern Idaho do not show similarly elevated mortality in mature stands.