低温和诱导时间对小麦小孢子培养的影响

为了优化小麦小孢子培养体系,研究了低温处理花药、幼穗及诱导培养时间对小孢子形态、胚胎发生和植株再生的影响。结果表明,经低温处理后,有活力的小孢子体积增大、形态变化明显,据其液泡大小、数目及其与细胞核位置的关系,可分为4种类型。低温处理可显著提高胚状体数和绿苗数,其中处理花药3 d、幼穗7 d的效果最好,平均每皿的绿苗数分别为59株和57株。胚状体发育速度显著影响绿苗分化能力,小孢子诱导培养28-29 d、直径约2 mm的胚状体数目较少,但绿苗分化率高;诱导培养32-38 d、直径为2 mm的胚状体数目达到了峰值,而绿苗分化率极低。在小麦小孢子培养中,应选择诱导28-29 d、直径2 mm的胚状体进行分化培养。     

赤霉病菌特异抗体-防御素融合蛋白的表达和功能鉴定

选择对赤霉病菌具有高度特异性和亲合力的单链抗体,与植物防御素构建成融合蛋白基因,将融合蛋白及防御素基因分别与细菌表达载体pGEX和植物表达载体pMBL4构建成重组载体,在细菌中经IPTG诱导表达GST融合蛋白,亲和层析纯化蛋白;经根癌农杆菌介导的真空渗入法在烟草叶片中瞬间表达,提取叶片总蛋白用于鉴定。抑菌活性分析表明,烟草叶片瞬间表达的防御素和抗体-防御素融合蛋白,均对赤霉病菌有强烈的抑菌作用;从细菌中纯化的防御素蛋白,也能有效抑制赤霉病菌生长。说明赤霉病菌抗体-防御素融合蛋白基因,可作为抗赤霉病资源,用于改良植物的抗赤霉病性。     

中国叩甲科昆虫属的分布类型与特征分析

通过统计中国科学院动物研究所标本馆、中山大学生物防治国家重点实验室昆虫标本室、华中农业大学昆虫标本室和深圳职业技术学院生物应用工程系昆虫标本室叩甲科昆虫定名标本和相关资料,分析了中国叩甲科昆虫属的区系分布特点。结果表明:除异角叩甲亚科Pityobiinae在古北界没有分布外,其余11个亚科在东洋界和古北界均有分布;东洋界的特有属很多,达到64属,占到总数的一半(50%)。从各区的分布情况看,分布于华南区的属最多,达到100属,占总数的78.1%;华北区和青藏区分布的属最少,分别只有21和14个属,分别占总数的16.4%和10.9%。中国叩甲科昆虫在中国动物地理区划中共有37个分布型,其中华南区的种类最多,达到37属,占总数的28.9%;其次为华南、西南、华中3区共有种类,为13属,占总数的10.2%;再其次为华南、华中2区共有种类,为8属,占总数的6.3%。     

植酸酶基因工程研究进展

植酸酶在动物、植物和微生物的磷代谢中起着重要作用,近年来随着真菌基因工程和植物基因工程研究的发展,已克隆出10多个真菌的植酸酶基因,并利用构建基因工程菌的方法,大大提高了微生物生产植酸酶的效率。通过转基因方法,目前已经成功地将几种真菌植酸酶基因转到水稻、玉米等作物中,通过特异表达提高了转基因作物自身植酸酶含量。本文从真菌基因工程和植物基因工程2个方面综述了植酸酶研究取得的进展,并比较和分析了植酸酶植物基因工程和真菌基因工程各自的优缺点。     

图像流式细胞仪在中华绒螯蟹血细胞分群及吞噬功能研究中的应用

为了更精确地对甲壳动物血细胞进行快速分类和功能分析,本研究以中华绒螯蟹为例,依据其血细胞内具有颗粒结构的特征,与常规显微观察对比,探讨了一种基于图像流式细胞仪的血细胞自动化分类新方法,并测量了体质量为(10±3) g的中华绒螯蟹的血细胞对直径1μm微球的吞噬情况。结果显示,两种方法都可将中华绒螯蟹血细胞分为4个类群。显微镜观察分类基于胞内可见颗粒,但由于缺乏精细的量化标准,批次样品的人工辨识结果波动较大;而图像流式方法以胞内全部颗粒结构为对象,利用高精度检测模块测量,结果更准确、客观,而且通量高、重复性好。测量结果显示,中华绒螯蟹血淋巴中无颗粒、小颗粒、中颗粒及大颗粒细胞占比分别为40.62%±2.65%、36.68%±6.84%、7.80%±1.16%和16.51%±5.60%,依据测量的颗粒特征区分的4个类群界限清晰,缺乏过渡样点,提示各类细胞之间可能没有相互转化。进一步的活体微球吞噬实验证实,中华绒螯蟹的4类血细胞都具有吞噬功能,并以无颗粒细胞为主要吞噬类群;吞噬微球的细胞比例在注射后6 h内呈钟型曲线变化,4 h可达峰值(5.69%±0.44%),表明中华绒螯蟹血细胞能高效清除血淋巴中的异物。研究表明,图像流式细胞仪适合于中华绒螯蟹的血细胞分类分析和功能研究,本研究结果为同类研究提供了重要参考,将有助于更全面了解甲壳动物血细胞的功能。     

Effects of site preparation and fertilizer application at planting on Eucalyptus tereticornis at Morogoro,Tanzania

Uniform nursery stocks of Eucalyptus tereticornis were used to establish an experiment factorially combining site preparation (three levels—bedding; disking; neither bedding nor disking), initial nitrogen applications (two levels—75 g ammonium sulphate per plant; 0 g ammonium sulphate per plant) and initial phosphorus applications (150 g triple super phosphate per plant; 0 g triple super phosphate per plant).Monitoring was carried out for 2 years by which time trees were up to 3.2 m tall and 5.0 cm in diameter at the root collar. Survival at 2 years was significantly higher among plants that had received nitrogen (65%) than those which had not (53%) and among plants on disked or bedded sites (62%) than those on unworked soil (53%). Monthly height increment was briefly, but significantly, increased in plants supplied with additional nitrogen and further enhanced if there was also additional phosphorus. Monthly diameter increment was, over most of the 1st year, significantly greater on sites prepared by bedding; significant increases also resulted, though for more restricted periods, from site preparation by disking and from provision of additional nitrogen. At the end of 1 year significantly higher concentrations of foliar potassium, suggesting enhanced drought hardiness, were detected for plants on bedded or disked ground. The concentration in these was 1%; on unworked ground the corresponding value was 0.8%. Combining bedding or disking with an initial application of nitrogen offset this effect.The short period over which growth benefits from the site preparation and nitrogen application treatments is noted but it is stressed that initial growth encouragement is desirable with Eucalyptus and that overall benefits, as an improved survial, may be long term. The value of phosphorus applications under the prevailing conditions is considered unproven.     

Effect of inhibitors of melanin biosynthesis on appressorial penetration and reductive reactions in Pyricularia oryzae and Pyricularia grisea

Appressorial penetration of onion epidermal walls by wild-type strains P-2 and O-42 of Pyricularia oryzae was more sensitive to fthalide (4,5,6,7-tetrachlorophthalide) than penetration by wildtype strain 4091-5-8 and black mutant strain BL-3 of Pyricularia grisea. Cerulenin completely blocked appressorial penetration by P. oryzae strain P-2 or P. grisea strain BL-3, but penetration capacity of these appressoria could be largely restored with 0.1 mM scytalone. Fthalide and tricyclazole inhibited the conversion of 1,3,6,8-tetrahydroxynaphthalene (1,3,6,8-THN) to 1,3,8-trihydroxynaphthalene (1,3,8-THN) and 1,8-dihydroxynaphthalene (1,8-DHN) as well as the conversion of scytalone to 1,8-DHN by cell-free extracts of P. oryzae or P. grisea. These inhibitors blocked the NADPH-dependent reductase reactions involved in the conversion of 1,3,6,8-THN to scytalone and the conversion of 1,3,8-THN to vermelone in the melanin biosynthetic pathway to 1,8-DHN. The two reductase reactions in extracts from P. oryzae P-2 were about 10 times as sensitive to fthalide or tricyclazole as those in extracts of P. grisea. Reductase activity with either 1,3,6,8-THN or 1,3,8-THN as substrates was present in only trace amounts in cell-free extracts of the buff mutant, P. oryzae P-2 m-1.     

Studies into the differential activity of the hydroxybenzonitrile herbicides: II. Uptake,movement and metabolism in two contrasting species

Uptake, movement, and metabolism of unformulated ioxynil and bromoxynil salts were investigated in Matricaria inodora and Viola arvensis. The morphology of these two species did not give rise to different spray retention and contact angles. After 7 days, uptake of [¹⁴C]ioxynil-Na reached 8.26% of applied ¹⁴C activity in M. inodora and 16.77% of that in V. arvensis compared with 1.54 and 3.83%, respectively, for [¹⁴C]bromoxynil-K. Over 98% of the ¹⁴C activity detected in the plant after 7 days remained in the treated leaves of V. arvensis following [¹⁴C]ioxynil-Na treatment. However, 8.7% of the ¹⁴C activity detected in [¹⁴C]ioxynil-Na-treated M. inodora was recovered from the apex and developing leaves reflecting a greater translocation. [¹⁴C]Bromoxynil-K was more mobile in both species and after 7 days 87.5 and 91.39% were detected in the treated leaves of M. inodora and V. arvensis, respectively. In both species the majority of translocated ¹⁴C activity was recovered from the apex and developing leaves. Up to 20% of the applied [¹⁴C]ioxynil-Na and [¹⁴C]bromoxynil-K was not detected within the treated plant. Extraction of treated plants revealed no detectable metabolic breakdown of ioxynil-Na to halogenated derivatives in either species. However, metabolic breakdown of bromoxynil-K was apparent in V. arvensis. No significant root exudation was detected when [¹⁴C]ioxynil-Na and [¹⁴C]bromoxynil-K were applied to hydroponically grown S. media and V. arvensis. Losses of ¹⁴C activity were due to herbicide volatility or degradation to volatile products on the leaf surface.     

苏云金芽胞杆菌CY1菌株的cry基因分析

通过醋酸钠-抗生素筛选法从河北省土壤中筛选得到了苏云金芽孢杆菌新菌株CY1。镜检可观察到伴孢晶体呈小菱形。SDS-PAGE分析结果表明菌株CY1表达的晶体蛋白分子量为130 kDa。利用25对通用引物检测cry1,cry1I,cry7,cry2,cry3,cry4/cry10,cry5,cry6,cry7,cry8,cry9,cry11,cry13/14,cry16/17,cry18,cry19,cry21,cry22,cry24/25,cry26/28,cry27/29,cry30,cry32,cry34/35,cry40基因,仅从CY1菌株中检测出cry7基因,该cry7基因N末端的1 215 bp片段所编码的氨基酸序列与已发表的Cry7Ab1(Acc No.:U04367)序列同源性达99%,仅有2个氨基酸的差异。用3对cry7Ab特异引物检测,进一步证实该菌株携带有cry7Ab基因。     

藏东南高寒土坡面细沟水流输沙能力变化特征

为探究高寒土坡面细沟侵蚀过程机理,建立科学的坡面细沟输沙能力因子模型,服务高寒坡面水土流失治理工作,以藏东南高寒土壤为研究对象,采用室内径流放水冲刷试验,探讨不同流量和坡度条件下细沟水流输沙能力特征。结果表明：（1）高寒土坡面细沟水流输沙能力的临界坡长随输沙能力的增加而变短,范围为3.27~8.31 m;坡度在15°~25°时,临界坡长大约稳定在5.0 m;（2）不同坡度下,高寒土细沟水流输沙能力与流量表现为明显的线性正相关关系（T_c=Aq）;小坡度的输沙能力受流量的影响程度大于大坡度;（3）高寒土细沟水流输沙能力与坡度可以用指数方程较好地表示（T_c=-ae^（-S/b）+c）,输沙能力随坡度的增大先快速增大,后逐渐过渡到平稳,当坡度达到15°时增幅平缓;（4）坡面细沟水流输沙能力可以用二元幂函数方程T_c=1697.83S^0.491q^1.043表示。通过本模型与其他模型的比较分析,ANSWERS模型在计算高寒土的坡面细沟水流输沙能力还值得商榷,本试验模型、Lei模型与Gao模型均能较好地模拟高寒区细沟侵蚀输沙能力。