Selection, regeneration and protein profile characteristics of NaCl-tolerant callus ofRobinia pseudoacacia L.

Changes in callus growth, differentiation potency and protein profile induced by salt stress were investigated in the calli induced from a hypocotyl ofRobinia pseudoacacia L. The NaCl treatment evidently obstructed the callus growth and the influences seemed to continue to be present in the selected calli even after they were restored in NaCl-free medium that could partially recover the callus growth, and some polypeptides were found to restore. The calli selected and restored intermittently every two weeks for one year completely lost their differentiation potency. The NaCl-tolerance test of the regenerated plantlets revealed that part of the plantlets that directly regenerated from the selected NaCl-tolerant calli were able to survive in 0.2 M NaCl, while most of the plantlets regenerated from the NaCl-nonselected calli or from restored calli were obviously damaged by this treatment. The SDS-PAGE assay indicated that a 54 kDa polypeptide increased distinctly to a very high level while a 41 kDa polypeptide decreased to a lower level in almost all the NaCl-tolerant plantlets. This observation led to the suggestion that the changes may play an adaptive role in allowing plantlets to survive under saline conditions.     

Plantlets regeneration from root explants ofRobinia pseudoacacia L. by suspension culture

A rapid and efficient method for the regeneration of plantlets from root explants ofRobinia pseudoacacia L. by suspension culture was established. The roots taken from aseptically grown 15-day-old seedlings were used as explants. It was determined that photoperiodicity was necessary for root proliferation, and that the promotive effect of IAA (3-indoleacetic acid) on root proliferation was better than that of IBA (3-indolebutyric acid). The roots cultured in 1/2 MS liquid medium containing 3 μM IAA and 1% sucrose at 25°C under 16-hour photoperiod with 50 μmol m⁻²s⁻¹ PPFD (photosynthetic photon flux density) shaking at 100 times/min reciprocally showed high efficiency for root proliferation. BAP (6-benzylaminopurine) was found to be essential to induce adventitious shoots from the roots, and the roots cultured in the medium supplied with 3 μM BAP combined with 1–6 μM IAA for 3 weeks under the same conditions as in the root proliferation period were most suitable for adventitious shoot inducement.     

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.     

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.     

Propagation methods for black locust (Robinia pseudoacacia L.) improvement 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. Black locust plants are commonly produced by two methods, by seed and by root cuttings. Tissue culture propagation can be considered as a relatively new method. Growing trees from seed is a relatively simple method for reliably producing seedlings on a large scale under a variety of circumstances. Mechanization of the method is easy and the production cost is relatively low. Propagation from root cuttings and tissue culture are valuable for reproduction of superior individuals or varieties. By applying these methods, superior traits of the selected trees can be preserved in the clones. Recent experiments demonstrated that micropropagated trees could be successfully transplanted into soil, hardened and grown in the field. Biography: Karoly Redei (1952), male, doctor, deputy director general of Hungarian Forest Research Institute H-1023 Budapest, Frankel Leó Str. 42 44, HUNGARY. Responsible editor: Chal Ruihai     

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.     

匈牙利刺槐（（Robinia pseudoacacia L.）林的经营研究

匈牙利刺槐（（Robinia pseudoacacia L.)是在17世纪初从北美引入到欧洲的第个森林树种。在匈牙利，刺槐是非常重要的速生树种，为经济和生态目的，可以成功营造刺槐人工林。结合该树种的生长特点选择营林措施，可以最大限度的提高刺槐人工林的成活率和生产力。本文根据林分结构和森林产量长期实验林的研究成果，提出了刺槐林的抚育作业次序。制定好的营林计划和模式将会培育出效益刺槐林，并能使土地经营者更好地接受该树种。同时，刺槐也是一个非常有用的能源生产树种，本文展示了对这方面的研究成果。图2表1参8。     

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.     

Investigation of anti-salt stress on tetraploid Robinia pseudoacacia

Tetraploid Robinia pseudoacacia was used as a main test material and diploid R. pseudoacacia was used as the control. The indices of shape, physiology and biochemistry, photosynthesis and anatomic structure of the young plants were investigated under salt stress (NaCl and Na₂SO₄). The treatment time was 30 d with an interval time of 7 d. Before and after treatment, the indices were measured. Results show that: 1) the growth of diploid R. pseudoacacia inhibited an evident symptom of salt damage and the leaf moisture content was lower under salt stress than that of control. But the tetraploid R. pseudoacacia was contrary. 2) The relative electric conductivity and proline (Pro) of tetraploid R. pseudoacacia increased slightly and had no significant difference compared with its control, which was contrary to diploid R. pseudoacacia. At the same time, three protective enzymes including perocidase (POD), superoxide (SOD) and catalase (CAT) kept higher activities at a post stage of salt stress to tetraploid R. pseudoacacia, which enhanced its anti-salt characteristics. Diploid R. pseudoacacia was sensitive to salt and had contrary information. 3) Salt stress had little influence to photosynthesis of tetraploid R. pseudoacacia. The net photosynthetic rate (P _n) and intercellular CO₂ concentration (C_i) had no significant changes, but those of diploid R. pseudoacacia decreased singificantly. 4) After salt stress, the anatomic structure of tetraploid R. pseudoacacia had a positive reaction, including the palisade parenchyma of diachyma, was prolonged and arranged more tightly. The spongy parenchyma was shrunk and was arranged tightly, which was contrary with diploid R. pseudoacacia. These data demonstrate that tetraploid R. pseudoacacia had superior anti-salt performance. __________ Translated from Journal of Plant Ecology, 2008, 32(3): 654–663 [译自: 植物生态学报]     

Preliminary studies on establishment of genetic transformation system in loblolly pine

A transformation system was established for loblolly pine (Pinus taeda L.) mature zygotic embryos usingAgrobacterium tumefaciens. The gene coding for the β-glucuronidase (GUS) gene was introduced into loblolly pine tissues and its transient expression was detected with histochemical staining. The influences of different genotypes.Agrobacterum concentrations. and cocultivation time on GUS expression and kanamycin resistant callus and shoot regeneration were investigated. The results showed that the highest GUS expression frequency (16.3%) and shoot regeneration frequency (78%) were obtained from genotype 9–1003 withAgrobacterium concentration decreased 9 times and cocultivation time of 56 hours respectively. GUS expression was obtained in all genotypes tested. The successful expression of the GUS gene in different genotypes suggested that it will be a useful transformation system for loblolly pine. (Responsible Editor: Chai Ruihai)     

Thermogenetic curves and thermokinetics of seed germination ofRobinia pseudoacaia

Seed germination process has closely relation with material transformation and energy exchange within the seed. Study on its thermal effect is important for understanding the mechanism and the influencing factors of the seed germination. The thermogenetic curves of seed germination ofRobinia pseudoacacia was measured by a new-type conductive microcalorimeter made in Wuhan University. The relationship was analyzed between the germination thermogenetic regulation and seed germination physiology. The thermogentic curves were further analyzed by thermokinetic theory to obtain the dynamic parameters and the thermokinetic model on seed germination ofRobinia pseudoacacia. The relationship of the thermogenetic power (μ w) and the germination time(h) of the germination process of 20 grainsRobinia pseudocacia seeds at 25°C wasP=208.77/[0.1937+0.8063exp(−0.06563t)] Foundation item: This project is supported by National Nature Science Foundation (30070200). Biography: Hu Yun-chu (1960-), male, associated Prof. in Central South Forestry University, Zhuzhou 412006, Hunan, P. R. China. Responsible editor: Song Funan     

Variation in germination character of Robinia pseudoacacia L. (Leguminosae) seeds at individual tree level

To demonstrate the seed dormancy and germination characters of Robinia pseudoacacia L., an exotic tall tree species in Japan, we applied scarification, cold stratification, diurnal thermal regime, heat shock, and/or winter weathering treatments to the seeds. These characters differed markedly among three seed sources (Trees K, B, and I). Scarification revealed that most seeds from Tree K showed physical dormancy, whereas those from Tree B had no physical dormancy. The seeds from Tree I showed weak physical dormancy so that the seeds eventually germinate without any treatments as time goes by. The physical dormancy in Tree K was broken by a long, high heat shock treatment. The results imply that seeds from Tree K respond to fire in natural conditions. Seeds from Tree I responded to a wide range of thermal regimes, except for long, high heat shock. In contrast, most seeds from Tree B absorbed water during cold stratification and some germinated. However, many seeds from Tree B died in the soil during the winter, presumably because of microbial and fungal infections. Seeds from Trees I and B acquired physical dormancy by weathering during the winter, implying that seedpods that remain in the crown function as an aerial seed bank. R. pseudoacacia may be able to produce various levels in dormancy with respect to the winter condition.     

Agrobacterium-mediated transformation of lombardy poplar (Populus nigra L. var.italica Koehne) using stem segments

Genetically transformed lombardy poplar (Populus nigra L. var.italica Koehne) plants were regenerated by co-cultivation of stem segments withAgrobacterium tumefaciens strain LBA4404 that harbored a binary vector (pBI121) which included genes for β-glucuronidase (GUS) and neomycin phosphotransferase. Successful transformation was confirmed by the ability of stem segments to produce calli in the presence of kanamycin, histochemical and fluorometric assays of GUS activity in plant tissues, and Southern blot analysis.     

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.     

A model for vertical distribution of fine roots in Robinia pseudoacacia plantations on the Loess Plateau

Based on a detailed investigation of vertical distributions of fine roots in Robinia pseudoacacia plantations at the Ansai Soil and Water Conservation Station, Shaanxi Province, a model was developed for the deep distribution of fine roots of R. pseudoacacia, which reflects the growth of fine roots affected by the mixed process of infiltration water and deep soil water. The maximum depth of the distribution h _max and the depth of the highest fine root density (FRD) h _p were determined and the maximum depth of infiltration water supplied for fine root growth h _q could also be calculated, h _q was considered as the approximate boundary between infiltration water and deep soil water in support of the growth of fine roots. According to the model, the soil water of R. pseudoacacia woodland in the profile could be classified into three layers: the first layer from the soil surface to h _p was the active water exchange layer, very much affected by precipitation; the second was the soil water attenuation layer, between h _p and h _q and largely affected by the vertical distribution of fine roots; the third was the relatively stable soil water layer below h _q, below which soil water did not change much. The percentage of infiltration water supplied for the growth of fine roots reached a level of 88.32% on the shaded slopes and 85.21% on sunny slopes. This indicated infiltration of precipitation played a crucial role in the growth of R. pseudoacacia in the gully region of the Loess Plateau. The research of interaction between the distribution of fine roots and soil water in the profile will help to explain the reasons for the complete drying out of soils and provide a theoretical basis for continuing the policy of matching tree species with sites on the Loess Plateau. Translated from Scientia Silvae Sinicae, 2006, 42(6): 40–48 [译自: 林业科学]     

匈牙利刺槐(Robinia pseudoacacia L.)改良的繁殖方法(英文)

在匈牙利,匈牙利刺槐(Robinia pseudoacacia L.)是重要的成林树种之一,且该树种的重要性在世界其它国家也日益增加。该树种的生产通常有两种方法:种子和根插条。组织培养对该种来说是一种相对较新的方法。对于在各种环境条件下大规模地生产苗木来说,种子栽培是相对简单的方法,机械化方法比较容易且生产成本相对较低。根插条和组织培养对于优势个体或品种繁殖是有价值的。利用这些方法,所选出的树木的优势品质可以在无性系得以保存。近期实验表明,微繁殖树木可以成功地移植到土壤中,在野外生长。     

区域化试验中饲料型四倍体刺槐生物量比较

通过对2年生3个四倍体刺槐无性系和1个普通无性系的生物量调查和对比，发现四倍体无性系对西北环境的适应性较好，其生物量均超过了当地普通刺槐，这表明，其作为再生型土本饲料在生态条件较差的西北地区有一定的开发潜力，由无性系在不同立地条件下比较发现，土壤因子对刺槐生长有较大影响，其中较为显著的是水分供给条件。     

刺槐高质量基因组DNA提取方法的研究

高效、稳定的基因组DNA提取是进行分子标记辅助育种研究的关键技术环节之一。以刺槐叶片为材料,将传统的CTAB法进行优化改良,结合组织研磨棒液氮研磨建立刺槐基因组DNA的提取方法。将提取获得的基因组DNA进行SSR分子标记技术研究,获得的条带清晰、稳定．表明提取获得的基因组DNA能很好的满足SSR分子标记的需要。为刺槐遗...     

刺槐种子中凝集素的纯化及性质研究

从刺槐(RobiniapseudoacaciaL.)的种子中分离纯化出2种凝集素RPsAⅠ和RPsAⅡ。种子经磨碎、过滤、离心、硫酸铵分级、透析、Sepharose4B离子交换和SephadexG 150分子筛层析,即可获得在PAGE和SDS PAGE上均呈现单一蛋白染色带的凝集素纯品,分子筛层析测得RPsAⅠ和RPsAⅡ的分子量分别为132000和130000,SDS PAGE测得亚基分子量分别为38000和31000。当2种凝集素的质量浓度分别达到7 80μg/mL和15 60μg/mL时能凝集兔血红细胞,凝集活性均可被L 阿拉伯糖、D 果糖、D 半乳糖、麦芽糖抑制;热稳定性实验结果表明2种凝集素对热处理均具有一定的稳定性;当pH>9和pH<5时,它们的活性都明显降低。