美味牛肝菌菌塘中菌根促生菌的筛选与鉴定

为了获得黑松Pinus thunbergii-美味牛肝菌Boletus edulis（Be）菌塘中的菌根促生菌（mycorrhiza helper bacteria,MHB）,为美味牛肝菌的人工培育提供理论基础,通过比较野生牛肝菌菌塘细菌胞外代谢产物对菌丝生长的促进作用以及细菌+Be真菌双接处理黑松幼苗对其苗高、地径、茎根比、干重、根侵染率的影响,筛选出菌根促生菌。结果表明,菌株B2和K2代谢产物可以显著促进Be菌丝生长,盆栽实验结果表明双接种Be+B2、Be+K2的黑松苗,苗高分别增长了17.71%和16.82%,地径增长了41.65%和35.63%,干重提高了44.71%和48.78%,根侵染率提高了23.20%和21.50%,茎根比下降了46.41%和35.40%。可见菌株B2和K2是黑松-美味牛肝菌的菌根促生菌。通过细菌的形态、生化特性、16S rDNA序列测定,菌株B2和K2均鉴定为蜡样芽孢杆菌Bacillus cereus。蜡样芽孢杆菌有望进一步开发为美味牛肝菌的菌塘生物肥料。     

有益芽孢杆菌受体菌研究

采用酸性茚三酮法测定了30株有益芽胞杆菌的赖氨酸产量,然后在不同的溶菌酶浓度下,对赖氨酸产量超过0．07g/L的21株菌进行原生质体转化质粒pUB110,测定原生质体形成率、原生质体再生率及转化频率,结果6103,6104,6120,6129四株菌的转化频率较高。最后,采用经典遗传学方法选育AEC抗性突变株,使赖氨酸积累提高。其中,B.licheniformis 6104诱变菌株610401能积累赖氨酸2．91g/L ,比出发菌株提高了17倍左右,转化率也提高了一个数量级。通过质粒的再转化试验及传代稳定性试验,进一步证实B.licheniformis 6104及其 突变菌株610401是较好的受体菌,尤其是用于赖氨酸合成酶基因的表达。     

芽孢杆菌原生质体的形成和质粒转化的研究

测定了芽孢杆菌属中21个种、68个菌株的原生质体形成率。在高渗缓冲液(SMMP 或SMN)中,原生质体形成率在90％以上的有37株。降低高渗缓冲液中钠离子浓度,有利于原生质体的形成。用质粒(pUB 110或pC194) DNA对16株芽孢杆菌的原生质体进行了转化试验。转化成功的共8株：纳豆芽孢杆菌AS 1.107、AS 1.921、幼虫芽孢杆菌AS 1．430、球芽孢杆菌AS 1.1362、迟缓芽孢杆菌#50、苏云金芽孢杆菌松蠋亚种AS 1．294、地衣芽孢杆菌# 18和坚强芽孢杆菌#28。原生质体在DM3再生培养基上的再生率分别为0．1％一19．2％,转化效率分别为1．4×102一1．0×105转化子／μgDNA。转化效率低或未转化成功的菌株,其原生质体的再生率一般都很低或不能再生,有的菌株在形成原生质体后发生自溶。     

高效解硅菌的分离鉴定及其促生能力

【背景】硅具有提高作物抗逆性、提高产量及改善品质的能力而被广泛报道,但传统施肥对硅肥的忽略使耕地土壤有效硅含量不断降低,因此建立高效提高土壤有效硅的方式尤为重要。【目的】土壤中有效硅的含量影响作物对硅的吸收利用,通过施用具有解硅能力的土壤微生物实现土壤有效硅的持续活化,不仅对作物产量和品质有重要意义,同时也可减少化肥的施用。【方法】利用选择培养基进行土壤解硅菌的富集、分离纯化,并测定其解硅能力、耐盐碱性、耐低氧性、解磷、解钾、解纤维素能力;利用孔雀绿染色及沙黄溶液复染法测定高效解硅菌芽孢形成率,最后通过盆栽试验探究高效解硅菌株对玉米的促生作用。【结果】筛选获得2株高效解硅菌,分别为菌株MB22及MB35-5,16S rRNA基因序列测定鉴定为Priestia aryabhattai。培养液有效硅浓度分别为空白处理的1.5倍和1.7倍,为对照菌株胶质类芽孢杆菌Paenibacillus mucilaginosus3016处理的1.1倍和1.2倍。2株高效解硅菌表现出较好的耐盐碱性,MB35-5功能最强,在pH 10.0、10% NaCl、10% KNO₃条件下培养液有效硅浓度为0.28-0.37 mmol/L。MB22和MB35-5产芽孢率分别为68%和55%,分别为对照菌株3016的1.69倍和1.37倍。MB35-5对磷、钾和纤维素都有一定的活化利用能力。盆栽试验表明,2株菌均显著提升了玉米株高及根系干重,其中MB35-5促生效果最好,平均株高是空白处理的1.39倍,是对照菌株3016处理的1.14倍,根系干重为空白处理的1.37倍,是对照菌株3016处理的1.24倍。【结论】筛选得到2株高效解硅菌MB22和MB35-5,其中高芽孢率的MB35-5还具有解磷、解钾、降解纤维素和促生功能,同时具备耐盐碱、耐低氧等抗逆性,表明该菌株具有进一步试验和研发的潜势。     

四株高产磷脂酶C新菌株的鉴定和分类

在前面研究高产磷脂酶C(PLC)菌株筛选及其抗血小板功能的基础上,对新筛选的四株高产PLC的754-1、779、970和1107菌株进行了鉴定和分类。通过形态特征、培养特征、生理生化特征以及16S rRNA序列测定及其同源性分析,证实754-1菌株与Bacillus cereus基本一致,因此将754-1菌株分类属于蜡状芽孢杆菌,命名为蜡状芽孢杆菌深圳株754-1,B．cereus shenzhen 754-1 strain。而779、970和1107三株菌则与Bacillus mycoides基本一致,因而将这三株菌分类属于蕈状芽孢杆菌,分别命名为蕈状芽孢杆菌深圳株779,B．mycoides shenzhen 779 strain,草状芽孢杆菌深圳株970,B．mycoides shenzhen 970 strain,和蕈状芽孢杆菌深圳株1107,B．mycoides shenzhen 1107 strain。这将为进一步利用这些菌株及其PLC打下坚实的基础。     

利用原生质体融合技术选育防治植物病虫害的基因重组菌株

本试验选用抗菌蛋白产生菌枯草芽孢杆菌(B.subtilis) TG26和晶体蛋白产生菌苏云金芽孢杆菌(B.thuringiesis subsp,pacifeus) AS1.904的营养缺陷型衍生株,在聚乙二醇的诱导下进行原生质体种间融合,获得了表现双亲遗传性状的种间融合菌株。融合率为7.52×10~6,融合子经传代10次,稳定率为19.5%。融合菌株的菌落和细胞形态与亲本株明显不同。通过SDS-聚丙烯酰胺凝胶电泳检测,融合菌株表达了亲本的抗菌蛋白和毒素蛋白。抑菌杀虫试验表明,融合重组菌株具有抑制多种植物病原菌和毒杀鳞翅目幼虫的能力。     

整肠生菌纯培养物的环境动力学研究

采用振荡培养研究了温度、PH值和不同培养基的环境因素对整肠生菌（地衣芽孢杆菌）消长的影响．通过对整肠生菌的动力学基础数据按计算作图的动力学研究结果表明：整肠生菌最适生长温度在35～37℃时，活菌数15─17.2×10~7／ml,是30℃和40℃时的1.26倍和1.2倍．最适PH值在5.5～7.5时,活菌数为4.15─7.55×10~8／ml,是PH值4.5和9.5时的13.l倍和13.2倍．ABC三种培养基的结果表明,培养基C最适合地衣芽孢杆菌生长，菌数最高，为69.05×10~8／ml．对数期的世代时间也最短，G=20min,比AB号培养基中培养时缩短2─6min．生长比速度（μ=0.035)最大，是AB培养基培养时的1.27倍．残废率较低,5.2％．环境因素与整肠生菌消长的这种动力学关系，为利用地衣芽孢杆菌发酵生产“整肠生”制剂的调控以及该菌剂的治疗机制方面研究提供了科学依据．     

有益芽胞杆菌受体菌研究

采用酸性茚三酮法测定了30株有益芽胞杆菌的赖氨酸产量,然后在不同的溶菌酶浓度下,对赖氨酸产量超过0.07g/L的21株菌进行原生质体转化质粒pUB110,测定原生质体形成率、原生质体再生率及转化频率,结果6103,6104,6120,6129四株菌的转化频率较高。然后,采用经典遗传学方法选育AEC抗性突变株,使赖氨酸积累提高。其中,B. licheniformis 6104诱变菌株610401能积累赖氨酸2.91 g/L,比出发菌株提高了17倍左右,转化率也提高了一个数量级。通过质粒的再转化试验及传代稳定性试验,进一步证实B.licheniformis 6104及其突变菌株610401是较好的受体菌,尤其是用于赖氨酸合成酶基因的表达。     

新组合菌系SCB329-SCB933利用L-山梨糖发酵产生维生素C前体——2-酮基-L-古龙酸的研究 Ⅰ.新组合菌系SCB329-SCB933的生物学特性

采用紫外照射、化学诱变和原生质融合等方法选育到一株性状更优良的突变株SCB329,并与新筛选的一株芽孢杆菌SCB933搭配组成新的组合菌系。产酸小菌SCB329与其亲本菌株氧化葡萄糖酸杆菌性状相似。伴生大菌SCB933属苏芸金芽孢杆菌（B.thuringiensis）。新组合菌系利用L-山梨糖的发酵液提取后经纸层析,元素分析和红外吸收光谱等项鉴定,其发酵产物确系2-酮基-L-古龙酸,对新组合菌系的生物学特性也进行了研究。     

一株芽孢杆菌在维生素C二步发酵中对小菌的促进作用

从土壤中分离到1株能更好促使小菌生长和产酸的芽孢杆菌B601,作为伴生菌与巨大芽孢杆菌相比,在生长过程中,发酵液中B601活菌数小于巨大芽孢杆菌,而其芽孢数则多于巨大芽孢杆菌。对B601组成菌系的发酵条件进行优化,得到如下结果：100g／L L-山梨糖、6g／L尿素、10g/L玉米浆、培养温度30℃和发酵周期44h。与巨大芽孢杆菌组成菌系相比其底物,L-山梨糖质量浓度提高了25％,尿素下降了50％．玉米浆质量浓度下降了33％,温度提高了2℃,发酵周期缩短了4h。结果表明：B601作为伴生菌,与巨大芽孢杆菌相比,该菌株明显提高了发酵效率。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.