园林苗木穿孔病发病规律及综合防治技术

穿孔病是园林苗木的主要病害之一,大多数核果类苗木都是易感病品种,此病主要为害叶片,也为害嫩梢、枝干和果实,防治难度比较大。笔者借助显微镜,利用观察法对田间病菌孢子传播情况和田间病害发生情况进行测报分析,取得了测报结果,总结出了田间发病的始期、盛期和末期,制定了一套综合防治方法,并用于田间防治,取得了良好的防治效果。     

Development and Identification of Introgression Lines from Cross of Oryza sativa and Oryza minuta

Introgression line population is effectively used in mapping quantitative trait loci(QTLs),identifying favorable genes,discovering hidden genetic variation,evaluating the action or interaction of QTLs in multiple conditions and providing the favorable experimental materials for plant breeding and genetic research.In this study,an advanced backcross and consecutive selfing strategy was used to develop introgression lines(ILs),which derived from an accession of Oryza minuta(accession No.101133) with BBCC genome,as the donor,and an elite indica cultivar IR24(O.sativa),as the recipient.Introgression segments from O.minuta were screened using 164 polymorphic simple sequence repeat(SSR) markers in the genome of each IL.Introgressed segments carried by 131 ILs covered the whole O.sativa genome.The average number of homozygous O.minuta segments per introgression line was about 9.99.The average length of introgressed segments was approximate 14.78 cM,and about 79.64% of these segments had sizes less than 20 cM.In the genome of each introgression line,the O.minuta chromosomal segments harbored chromosomal fragments of O.sativa ranging from 1.15% to 27.6%,with an overall average of 8.57%.At each locus,the ratio of substitution of O.minuta alleles had a range of 1.5% 25.2%,with an average of 8.3%.Based on the evaluation of the phenotype of these ILs,a wide range of alterations in morphological and yield-related traits were found.After inoculation,ILs 41,11 and 7 showed high resistance to bacterial blight,brown planthopper and whitebacked planthopper,respectively.These O.minuta-O.sativa ILs will serve as genetic materials for identifying and using favorable genes from O.minuta.     

Interleukin 1, a mediator of immunoadjuvant peptidoglycans

Bacterial peptidoglycans and the synthetic analog muramyl dipeptide possess various immunomodulating properties (adjuvant effect, increase of resistance to infectious agents and to tumor growth). They are able to induce B cell activation and to stimulate macrophages to produce monokines such as Interleukin 1 (IL 1). IL 1 plays an essential role in immune response. It promotes thymocytes maturation and Interleukin 2 secretion by antigen sensitive T cells, which in turn triggers regulatory T cells. Moreover, it is involved in the proliferation and differentiation of B cells.

There is a correlation between the immunoenhancing effect of PG of a definite structure and their ability to induce IL 1 secretion. Non-adjuvant PG were inactive. This suggests that one of the major mechanisms of action of adjuvant PG could be the stimulation of IL 1 synthesis.     

大黄鱼C型凝集素受体Clec4e的分子鉴定及凝集特性

C型凝集素受体（C-type lectin receptor，CTLR）是可以特异性结合糖类病原体相关分子模式（pathogen-associated molecular patterns，PAMPs）的模式识别受体（pattern recognition receptors，PPRs），在先天性免疫中发挥着重要作用。为了揭示硬骨鱼CTLR的生物学功能，本研究以从大黄鱼（Larimichthys crocea）转录组数据库中筛选出的一个CTLR基因—C型凝集素结构域家族4成员E基因（C-type lectin domain family 4 member E gene，Clec4e）为研究对象，研究其分子特征、表达分布和凝集特性。结果显示，LcClec4e cDNA全长1546 bp，开放阅读框（open reading frame，ORF）771 bp，编码254个氨基酸。LcClec4e的N端有一个跨膜区，无信号肽，C端含有一个糖识别结构域（carbohydrate recognition domain，CRD），其中含有糖结合位点EPN和WFD以及6个可形成二硫键的保守半胱氨酸。系统发育分析表明，LcClec4e与多种鲈形目鱼类Clec4e具有较近的亲缘关系。荧光定量PCR结果显示，LcClec4e在所检测的10种组织中呈组成型分布，且在肝脏中表达量最高；LcClec4e在来源于大黄鱼头肾组织的原代巨噬细胞、淋巴细胞和粒细胞中均有表达，且在巨噬细胞中表达量最高；经灭活溶藻弧菌刺激后，LcClec4e在3种免疫细胞中的表达均极显著上调。原核表达的重组LcClec4e胞外段（recombinant LcClec4e-extracellular domain，rLcClec4e-ex）具有Ca2+依赖性的凝集活性，可凝集小鼠、家兔的红细胞，以及嗜水气单胞菌、变形假单胞菌、溶藻弧菌和坎氏弧菌等4种水产常见的革兰氏阴性菌。D-葡萄糖、D-果糖、D-甘露糖、D-麦芽糖、α-乳糖和脂多糖均可抑制rLcClec4e-ex对大黄鱼重要病原菌变形假单胞菌的凝集作用，说明LcClec4e可能与变形假单胞菌表面的糖类物质结合。这些研究结果提示，LcClec4e可能作为一种PPR，通过结合病原菌表面的糖类PAMPs来识别病原，参与大黄鱼抗细菌感染的免疫防御。     

Bacterial community structure of nirK-bearing denitrifiers and the development of properties of soils in created mitigation wetlands

We investigated the abundance and genetic heterogeneity of bacterial nitrite reductase genes (nir) and soil structural properties in created and natural freshwater wetlands in the Virginia piedmont. Soil attributes included soil organic matter (SOM), total organic carbon (TOC), total nitrogen (TN), pH, gravimetric soil moisture (GSM), and bulk density (D_b). A subset of soil attributes were analyzed across the sites, using euclidean cluster analysis, resulting in three soil condition (SC) groups of increasing wetland soil development (i.e., SC1 < SC2 < SC3; less to more developed or matured) as measured by accumulation of TOC, TN, the increase of GSM, and the decrease of D_b. There were no difference found in the bacterial community diversity between the groups (p = 0.4). NirK gene copies detected ranged between 3.6 × 10⁴ and 3.4 × 10⁷ copies g⁻¹ soil and were significantly higher in the most developed soil group, SC3, than in the least developed soil group, SC1. However, the gene copies were lowest in SC2 that had a significantly higher soil pH (~6.6) than the other two SC groups (~5.3). The same pattern was found in denitrifying enzyme activity (DEA) on a companion study where DEA was found negatively correlated with soil pH. Gene fragments were amplified and products were screened by terminal restriction fragment length polymorphism (T-RFLP) analysis. Among 146 different T-RFs identified, fourteen were dominant and together made up more than 65% of all detected fragments. While SC groups did not relate to whole nirK communities, most soil properties that identified SC groups did significantly correlate to dominant members of the community.     

广西芒果细菌性坏死病病原菌鉴定

芒果Mangifera indica L.是我国著名的热带水果, 近年来细菌性病害发生严重。2020年－2021年对广西百色地区的芒果病害进行调查发现, 一种细菌性坏死病与细菌性黑斑病混合发生, 一般发病率30%~60%, 严重可达90%以上。本研究先后从该地区不同芒果坏死病组织中分离得到泛菌属的21株细菌。根据形态、生理生化特性、16S rDNA、fusA、gyrB、leuS、pyrG、rlpB和rpoB的多基因系统发育分析、致病性测定等方法将21个菌株分别鉴定为Pantoea vagans、P. anthophila、P. dispersa 和P. cypripedii。其中P. anthophila、P. dispersa 和P. cypripedii引起芒果细菌性坏死病是中国首次报道。     

Mechanism of systemic inflammatory response syndrome induced by EC DNA

AIM: To investigate whether the bacterial DNA participates in SIRS and its possible mechanism. METHODS: Escherichia coli genomic DNA (EC DNA) was extracted and purified from Escherichia coli 25922 by alkaline lysis method. Mortality of mice challenged with EC DNA and the changes of TNF-α and IL-6 in rat serum were observed. ANA-1 cells were cultured in vitro, after the cells were stimulated by different concentrations of EC DNA and LPS, the level of TNF-α and IL-6 in supernatant were tested. Meanwhile,expression of TLR9 and TLR4 on cell surface was measured. Activation of NF-JB was also observed. RESULTS: The lethal effect of EC DNA on mice with an obvious dose-effect relationship was observed. The death happened within 24 hours. Calf thymus DNA and DNase I-treated EC DNA did not lead to mice to die. The changes of serum TNF-α and IL-6 in rats induced by EC DNA and LPS were similar, but TNF-α peak level of EC DNA group appeared 1 hour earlier than that of LPS group. In vitro, large amount of TNF-α and IL-6 were released from ANA-1 cells stimulated by EC DNA. High expression of TLR9 and TLR4 was observed on surfaces of THP-1 cells. In particularly, LPS induced strong activation of NFκB. The results suggested other pathway possibly took part in the signal transduction inducea by EC DNA. CONCLUSION: EC DNA has the abilities to lead to death of mice, andinduces serum TNF- αand IL- 6 level to increase in rats and ANA- 1 cells to release cytokines in vitro. High expression of TLR9 and TLR4, strong activation of NF- κB may be its importantmolecular mechanism, but other pathway probably exists to play an important role.     

盐酸小檗碱与左氧氟沙星联合抑制MRSA及其机制研究

养殖场抗生素滥用造成多种耐药性细菌的产生,从奶肉类制品中分离出耐甲氧西林金黄色葡萄球菌（MRSA）的案例逐年增长,对养殖业食品安全造成巨大威胁。本研究筛选出与左氧氟沙星具有联合抑制MRSA效果的天然产物盐酸小檗碱（BBR）,探究BBR与其对MRSA的协同抑菌效果和联合抑菌机制。结果表明,BBR能有效抑制MRSA生长,提高MRSA对左氧氟沙星的敏感度,两者联用后,MRSA对左氧氟沙星的MIC降为之前的1/16。协同作用机制主要为上调ribA及下调mec A和msc L表达水平,破坏细胞壁、增加细胞膜的通透性,从而达到协同抑菌的目的。本研究有助于降低畜牧养殖中抗生素的使用,为提高肉奶类食品安全奠定基础。     

双组分调控系统介导革兰阴性菌耐药的作用机制

菌群耐药已成为临床上亟待解决的关键问题,特别是革兰阴性菌引起的耐药现象尤为突出,给临床治疗带来了巨大的挑战,尽快阐明重要细菌复杂耐药表型的调控机制就显得尤为重要。双组分调控系统(two-component regulatory systems,TCS)存在于多种革兰阴性菌中,在细菌诸多生命活动中发挥关键作用,是细菌感知环境变化并产生相应调控的主要机制之一。TCS通常由两种蛋白组成,包括感受器蛋白(通常是组氨酸激酶)和反应调节蛋白(通常是转录因子),二者可通过磷酸化介导的协同作用,整合细菌周围的环境信号、调节细菌相关的基因表达及改变细菌的某些生理行为。近年来,探索细菌TCS介导的耐药性应答机制已成为一个新的研究热点。基于此,本文从TCS介导临床重要革兰阴性菌耐药的结构基础和作用机理等方面进行综述,以期增进对细菌TCS的全面认识,为今后临床上药物的科学研发提供新的思路和对策。     

芽孢杆菌产纤维素酶的研究

对芽孢杆菌 (Bacillussp.ZU 0 4 )产纤维素酶的工艺参数进行了优化 ,研究结果表明 :木糖渣和豆饼粉分别是该菌合成纤维素酶的适宜碳源和氮源 ,NaCl和KH2 PO4对纤维素酶的合成具有重要作用 ,其适宜质量用量分别为 0 .5%～ 1.0 %及 0 .1% ,麸皮的添加可明显提高发酵液中的酶活力 ;3L发酵罐中的适宜发酵条件为 :搅拌速度 30 0r/min ,通气量 0 .3L/(L·min) ,培养温度 37℃ ,中性和碱性纤维素酶活力分别达到了2 57.6和 12 5.6U/mL ,显示了良好的工业应用前景。