Factors involved in Agrobacterium tumefaciens-mediated gene transfer into Pinus nigra Arn. ssp. salzmannii (Dunal) Franco

Cotyledons from dissected sterile embryos of salgareño pine (Pinus nigra Arn. ssp. salzmannii (Dunal) Franco) were inoculated with different disarmed Agrobacterium tumefaciens strains harbouring the binary vector p35SGUSint. The transient expression of a β-glucuronidase gene (uidA) was studied, using a histochemical staining procedure. Nineteen days after inoculation, the activity of β-glucuronidase was detected in epidermal and subepidermal layers of cotyledonary explants. The EHA105 strain harbouring a disarmed agropine-type Ti-plasmid (pTiBO542) was the most effective for gene transfer of the uidA gene. The effects of exudates and extracts from 0-day-old embryos on induction of vir gene expression in A. tumefaciens were also examined. The results of this study showed that salgarño pine embryo exudates contain a substance(s) that induce vir gene expression, in similar way to that observed with 100 μM acetosyringone (AS).All these findings suggest that T-DNA processing and transfer might take place when Agrobacterium infects suitable tissues of salgareño pine.     

松南结缕草成熟胚愈伤组织的诱导和再生

松南结缕草成熟种子经灭菌和打破休眠后,接种于含有不同浓度2,4-D的MS和N6培养基上进行愈伤组织诱导。结果表明,结缕草种子的休眠主要在于破除种壳。2,4-D浓度在2~5 mg/L对愈伤组织诱导率无明显影响。基本培养基MS和N6诱导效果相近,初生愈伤组织为白色、半透明,质地松软、水浸状。继代培养基中添加一定浓度的甘露醇、麦芽糖和ABA以及延长继代时间有助于愈伤组织的胚性化。基本培养基MS更适于结缕草初生愈伤组织致密胚性化。胚性愈伤组织在MS添加0.1 mg/L 2,4-D培养基上即可分化出大量具有根和茎叶的再生植株。     

草地早熟禾新格莱德胚性愈伤组织原生质体培养及植株再生的研究

以草地早熟禾品种——新格莱德成熟种子为供试材料,在含3.0mg/L2,4-二氯苯氧乙酸(2,4-D)、0.5mg/L6-苄氨基嘌呤(6-BA)的MB₅培养基中进行胚性愈伤组织诱导的培养。并从生长了7～9个月的胚性愈伤组织中分离出原生质体,将该原生质体置于KM₈P培养基(含3.0mg/L2,4-D、0.5mg/L6-BA、100mg/L水解酪蛋白、100mg/L水解乳蛋白、1%蔗糖、0.4mol/L甘露醇)中进行了液体浅层培养。结果表明,新格莱德原生质体在上述KM₈P培养基中培养3d后出现第1次细胞分裂,2～3周后形成小细胞团,此时添加低渗培养液2～3次,小细胞团持续分裂并形成愈伤组织。当愈伤组织块长至3～5mm时,转入固体培养基MS+3.0mg/L2,4-D+0.5mg/L6-BA和MS+0.5mg/L萘乙酸(NAA)+5.0mg/L6-BA上进行培养,使其细胞增殖和分化,且逐步形成完整的植株。     

培养基质,赤霉素和硼对蕨菜孢子萌发成苗影响的研究

为了提高蕨菜孢子人工培养成苗效率,对４ 种培养基质和赤霉素（ＧＡ３）、硼３ 种使用水平的孢子萌发成苗效果进行了研究。结果表明,森林土、琼脂和蛭石间原叶体数无差异,但成苗率间差异显著;森林土成苗率最高,达到４２．８６％ ;砂基上孢子虽能萌发产生片状体,但不能发育到性器官形成期;赤霉素对孢子萌发生长具有显著的促进作用,在含有０．１５×１０－ ７ ｇ／Ｌ赤霉素培养基上成苗率较高;硼可提高配子体受精成苗,并以０．０５％ 硼酸处理效果最好;赤霉素与硼有显著的互作效应,含有０．１５×１０－ ７ ｇ／Ｌ赤霉素培养基上生长的原叶体,在性器官形成期喷施０．０５％ 硼酸,成苗率高达５１．６０％ 。     

无血清培养基IVD101和G1/G2在牛体细胞核移植中的应用研究

通过考察牛体细胞核移植重构胚在无血清培养基（IVD101、G1／G2）条件下培养的囊胚发育率及其质量,从而评估无血清培养基支持牛体细胞核移植重构胚体外发育的能力。采用IVD101和G1／G2对牛体细胞核移植胚胎进行体外培养,并以CR1aa＋5％FBS作为对照组。无血清培养基IVD101和G1／G2的囊胚发育率与对照组无显著性差异（41．2％±9．1％、42．2％±10．8％,48．0％±9．2％,P〉0．05）。通过囊胚差异染色和冷冻／解冻胚胎存活率分析胚胎质量,发现无血清培养基ICM／Total略低于对照组（31．8％±10．5％、29．5％±11．9％vs ．33．0％±14．8％）,但无显著性差异（P〉0．05）;3个组别的囊胚经程序化冷冻／解冻后无血清培养基的存活率（IVD101、G1／G2）略高于对照组,但差异不显著（84．8％、80．4％＇US．77．3％,P〉0．05）。结果证明无血清培养基（IVD101、G1／G2）可以支持体细胞核移植重构胚的体外发育,且其对程序化冷冻的耐受性与添加血清组（CR1aa＋5％FBS）相似。     

Mixed Forests as a Part of Sustainable Forestry in Southern Sweden

Abstract

We review and discuss the applicability of mixed stands in the context of sustainable forestry in southern Sweden. This is done from a silvicultural perspective regarding aspects such as regeneration, pre-commer-cial thinnings, production, economy, damage and policy implications. The paper is based on literature reviews and on studies performed in southern Sweden or under comparable conditions. After considering the underlying mechanisms, we find that it is possible to establish a mixed stand in the regeneration phase and sometimes even to a lower cost compared to monocultures. To keep the mixture, or to create it, with pre-commercial thinnings is, however, often more expensive. The reviewed studies, together with a new simulation, show that the effect on productivity of mixed stands compared to monocultures is minor. Some positive effects on damage (i.e., decreased levels) in mixed stands have been found. The economy and the policy implications of mixed stand management were found to be dependent on the specific stand and situations. A general finding was that research and knowledge of managing mixed stands, as compared to monocultures, are limited, which in turn could limit the applicability of mixed stand management.     

皇帝蕉(Musa AA)组织培养与快速繁殖技术研究

摘要：对皇帝蕉芽的消毒方法、外植体的诱导、幼芽的增殖培养、生根培养等进行了研究。结果表明：采用75%酒精60s--0.1%升汞12min（第一次8min 第二次4 min）的消毒方法,比较适宜于皇帝蕉外植体的消毒,易获得理想的无菌体系;在芽的诱导初期阶段,6-BA浓度采用5.0 mg/L,最适合皇帝蕉外植体的诱导;在幼芽的中高代增殖培养阶段,适宜采用MS 4.0mg/L6-BA 0.1mg/LNAA配方;在幼芽的高代增殖培养阶段,适宜采用MS 3.0mg/L6-BA 0.1mg/LNAA配方;在幼芽的生根阶段,适宜采用MS 2.5mg/LIBA 0.2mg/LNAA配方,幼苗生根快、根系壮实、发达,成活率高。     

Purified Culture Systems for Bovine Oviductal Stromal Cells

Isolated stromal cells from the ampullary and isthmic parts of bovine oviductal tissues were cultured in monolayer and spheroid (cell aggregate) systems. Prostaglandin F2α (PGF) plays a crucial role in oviductal contraction and is produced by oviductal epithelial cells in cattle. Since stromal cells of many organs produce PGF, PGF production by bovine oviductal stromal cells was investigated. After PGF synthesis was confirmed, the utility of isolation and culture methods for oviductal stromal cells was evaluated by PGF production in the present study. The homogeneity of the cells was > 99%. PGF production of the cells was increased by tumor necrosis factor-α. The stromal cells aggregated and formed a spheroid by the treatments with several reagents. PGF production was higher in the spheroid culture than in the monolayer culture. The isolation and culture methods described here will facilitate studies of the physiological function of bovine oviductal stromal cells.     

牛体外受精卵的二步法培养体系的研究

以CR1aa为基础培养液,采用二步法对牛体外受精卵进行体外培养,完善牛体外受精卵的培养体系.实验一:对照组连续7 d均为CR1aa 50 mL/L FBS培养,处理组前3 d为CR1aa 3 mg/mLBSA培养,后4 d换为CR1aa 50 mL/L FBS.处理组的囊胚率显著高于对照组,但卵裂率和囊胚孵化率无显著差异.实验二:对照组连续7 d均为CR1aa 50 mL/L FBS培养,处理组前3 d为CR1aa培养,后4 d换为CR1aa 50 mL/L FBS.处理组的卵裂率显著高于对照组,但囊胚率和囊胚孵化率差异不显著.实验三:对照组连续7 d均为CR1aa 50 mL/L FBS 0.1mmol/L GSH培养,处理组前3 d为CR1aa 0.1 mmol/L GSH培养,后4 d换为CR1aa 50 mL/L FBS 0.1 mmol/L GSH.处理组的卵裂率显著高于对照组,囊胚率极显著高于对照组,但囊胚孵化率差异不显著.结果表明,GSH与二步法培养系统结合相对于传统的一步法培养系统更适于牛体外受精卵的体外培养.     

Vernalization of immature embryos of winter wheat genotypes

Summary The effect of direct vernalization of immature embryos on flowering was studied in six winter wheat genotypes. Fourteen-, 17-, and 20-day-old embryos were excised and vernalized for 0–6 weeks on synthetic medium during a conditioning period. Percent germination of embryos was high (overall 96.1%), and free from genotypic effects. Genotypes differed for flowering in response to cold treatment of excised embryos. Embryo vernalization was as effective as or more than conventional vernalization (control, seedling vernalization for 6 weeks). Seventeen-day-old embryos were the most responsive to vernalization. With a 5-week vernalization of 17-day-old embryos, the percentage of plants anthesed was higher than those from 14-and 20-day-old embryos. For 17-day-old embryos vernalized for 5 weeks, the mean number of days from culture to anthesis was less than that of 6 week vernalization, less than that of 14- and 20-day-old embryos, and less than controls.Purdue Univ., Agronomy Dept., W. Lafayette, IN 47907, USA.