黑曲霉柠檬酸工业菌株原生质体制备与转化

黑曲霉是柠檬酸工业化生产的主要发酵菌株。尽管现代遗传操作技术在黑曲霉的实验室菌株、蛋白质生产菌株等的应用中获得了成功,但是柠檬酸工业菌株遗传转化异常困难,成为柠檬酸工业持续升级的重要限制因素。以柠檬酸工业生产实际应用的黑曲霉菌株为研究对象,对其原生质体的制备与再生以及PEG介导的转化等条件进行了细致优化。结果表明,柠檬酸高产工业菌株原生质体的制备需选取丰富培养基中培养48 h的年轻菌丝体,在1.5%裂解酶-0.5%蜗牛酶-0.2%溶菌酶的复合酶解体系下裂解2.5 h,原生质体的制备浓度可达106个/m L以上,原生质体再生效率可达90%以上。原生质体的浓度是原生质体-PEG介导转化方法的关键,当原生质体浓度达到106个/m L以上时,高产柠檬酸菌株的转化效率大幅提高。成功建立了由原生质体-PEG所介导的高产柠檬酸黑曲霉菌株的遗传转化体系。     

The circadian regulation of extracellular ATP

Extracellular ATP is a potent signaling molecule released from various cells throughout the body and is intimately involved in the pathophysiological functions of the nervous system and immune system by activating P2 purinergic receptors. Recent increasingly studies showed that extracellular ATP exhibits circadian oscillation with an approximately 24-h periodicity, which participates in regulatory pathways of central oscillator suprachiasmatic nucleus and peripheral oscillator bladder, respectively. Oscillators modulate the protein expression of ATP release channels and ectonucleotidase activity through clock genes; indeed, real-time alterations of ATP release and degradation determine outcomes of temporal character on extracellular ATP rhythm. The regulatory pathways on extracellular ATP rhythm are different in central and peripheral systems. In this review, we summarize the circadian rhythm of extracellular ATP and discuss several circadian regulatory pathways in different organs via ATP release and degradation, to provide a new understanding for purinergic signaling in the regulatory mechanism of circadian rhythm and a potential target to research the circadian regulation of extracellular ATP in other circadian oscillators.     

杨树胞外蛋白的提取分离及2-D电泳体系的建立

该研究以美洲黑杨杂种优良无性系NL895杨(Populus deltoides×Populus euramericana cv.)组培苗叶片和茎段为研究对象,对NL895杨叶片细胞壁蛋白(cell wall proteins,CWPs)和茎段质外体蛋白(apoplastic proteins,APPs)的提取、分离和双向电泳等技术进行了系统研究。结果表明:10g以上叶片、超声波破碎10min、CaCl2法提取的细胞壁蛋白效果较好,经G-6-PDH酶活性检测,提取的细胞壁蛋白胞质污染率较低;真空渗透法提取的茎段质外体蛋白胞质污染率较前者高,但在允许范围之内。TCA/丙酮沉淀法纯化提取的细胞壁蛋白、pH 4~7的24cm胶条、上样量为500μg的双向电泳体系,其蛋白电泳图谱中的斑点多而清晰,斑点数达550多个,是叶片细胞壁蛋白电泳分析较适合的体系;pH 3~10胶条对茎段质外体蛋白的电泳分离效果较好。该研究初步建立了杨树胞外蛋白的提取、分离及2-DE电泳体系,为木本植物胞外蛋白的研究奠定了基础。     

SGTA基因及其蛋白的生物信息学分析

[目的]对SGTA基因及其蛋白的结构和特征进行生物信息学分析,为研究SGTA与肿瘤形成和发展的相关性提供理论基础。[方法]运用生物信息学数据库和软件对SGTA基因的结构、单核苷酸多态性位点(SNP)、SGTA基因与其他基因的相互作用网络、SGTA蛋白的理化性质、二级结构、蛋白结构域、蛋白翻译后修饰、蛋白质之间相互作用网络进行分析。[结果]人SGTA基因有5种可变剪接产物,编码区存在78个SNP位点,其中错义突变31个,无义突变1个。人SGTA蛋白由313个氨基酸组成,是稳定性不高的亲水蛋白,α-螺旋是其主要二级结构元件,属于TRP超家族,预测有3个磷酸化激酶修饰位点和数个潜在泛素化修饰位点。与SGTA存在相互作用的基因和蛋白多数与维持体内蛋白质稳定的分子伴侣功能相关。[结论]SGTA基因及其蛋白的生物信息学分析为进一步实验研究其在肿瘤形成和发展中的地位及调控机制奠定了基础。     

利用宏基因组学技术分析煤层水中细菌多样性

煤层气是一种重要的能源资源,有生物成因和热成因两种类型。生物成因煤层气是在煤层微生物的作用下形成的。本研究通过向煤层气井中注入培养基,采用宏基因组学技术对煤层水中细菌菌群的组成多样性进行分析。结果表明,在培养基注入前后,煤层水中微生物均具有较丰富的多样性,注入前丛毛单胞菌科、甲基球菌科、假单胞菌科、鞘氨醇单胞菌科和嗜甲基菌科的微生物为优势菌种。注入3个月后,优势菌种主要为链球菌科、丛毛单胞菌科、嗜甲基菌科和红环菌科。注入6个月后,红环菌科和甲基球菌科的丰度最高。主成分分析显示,红环菌科、甲基球菌科、弯曲杆菌科、鞘氨醇单胞菌科、假单胞菌科、丛毛单胞菌科、嗜甲基菌科和链球菌科8个科的微生物对生物成气过程有重要影响。细菌菌群的ɑ多样性从chao1、ACE、Shannon、npS hannon和Simpson指数进行分析,结果显示,培养基的注入能显著增加物种的多样性。本研究结果为解析煤层生物成气过程中微生物的动态变化提供了依据。     

喀斯特峰丛洼地不同类型森林养分循环特征

以中国西南喀斯特峰丛洼地为研究区域用标准木法和收获法对人工林、次生林、原生林3个不同类型森林的6个代表性群落的生物量、营养元素生物循环量及循环特征进行了研究。结果表明:(1)不同类型森林群落乔木各器官的养分含量大小顺序为:叶枝根干,林下植被层和凋落物层的养分含量比较高,其含量普遍高于乔木层各组分,仅次于乔木叶片;各组分中营养元素以K、Ca最高,P、Mg最低;(2)3种类型森林间乔木层的养分积累量总规律表现为原生林(4540.30 kg/hm~2)次生林(2107.09 kg/hm~2)人工林(719.51 kg/hm~2),分别占林分养分积累量的88.30%、79.57%和62.60%;(3)3种类型森林生态系统养分总贮量相差不大,均主要集中在土壤层在各层分配格局有所差异;营养元素的年吸收量和年归还量均为次生林原生林人工林,年吸收量分别为:418.80、271.17和148.79 kg hm~(-2)a~(-1);年归还量分别为:182.98、111.43和43.37 kg hm_(-2)a~(-1);(4)不同类型森林养分利用系数总规律为人工林(0.35)次生林(0.20)原生林(0.10);循环系数则相反,为原生林(0.48)次生林(0.46)人工林(0.30);而周转时间为原生林(37.32)人工林(18.63)次生林(13.93)。喀斯特峰丛洼地土层薄,养分贮存能力差,森林养分循环能力相对较弱,沿着强、中、弱干扰递减梯度,3种类型森林养分利用效率和循环能力呈增长趋势。     

绿化植物的生活型对边坡植被物种多样性及护坡性能的影响

为了探讨绿化植物生活型构成对边坡植被物种多样性及护坡性能的影响, 揭示生活型构成-群落特征-物种多样性-生态系统功能间的内在联系, 借助三物种组装实验, 分别构建以草本、灌木或乔木为主体的草本型(HHX_i)、灌木型(SSX_i)、乔木型(AAX_i)或草-灌-乔混合型(HSA)配置模式的实验小区, 对实验区内边坡植被的群落特征、护坡性能进行持续4年的生态监测。结果表明: (1)边坡植被的物种丰富度与绿化植物生活型构成有关, AAX_i的物种丰富度总体高于其他模式, 呈AAX_i > SSX_i > HSA > HHX_i趋势。(2)不同配置模式边坡植被的群落盖度不一样, 年际间差异显著: 建坪初期(2010-2011年), HHX_i的群落盖度远高于其他模式, HSA次之, AAX_i最低; 2012-2013年间, HSA的群落盖度最高, HHX_i次之, AAX_i最低。(3)边坡植被的Shannon-Wiener指数、Pielou指数与绿化植物生活型构成有关, 其变化规律与群落盖度类似: 建坪初期, HHX_i的多样性水平远高于其他模式, 呈HHX_i > HSA > SSX_i > AAX_i趋势; 之后, 呈HSA > HHX_i > SSX_i > AAX_i趋势。(4)边坡植被的护坡性能与群落内的物种多样性密切相关: 多样性水平越高, 护坡性能越强。可见, 在生态护坡过程中, 绿化植物生活型构成对提高边坡植被物种多样性、改善护坡性能至关重要。     

Endophytic microbes <Emphasis Type="Italic">Bacillus</Emphasis> sp. LZR216-regulated root development is dependent on polar auxin transport in <Emphasis Type="Italic">Arabidopsis</Emphasis> seedlings

Key message

Endophytic microbes Bacillus sp. LZR216 isolated from Arabidopsis root promoted Arabidopsis seedlings growth. It may be achieved by promoting the lateral root growth and inhibiting the primary root elongation.

Abstract

Plant roots are colonized by an immense number of microbes, including epiphytic and endophytic microbes. It was found that they have the ability to promote plant growth and protect roots from biotic and abiotic stresses. But little is known about the mechanism of the endophytic microbes-regulated root development. We isolated and identified a Bacillus sp., named as LZR216, of endophytic bacteria from Arabidopsis root. By employing a sterile experimental system, we found that LZR216 promoted the Arabidopsis seedlings growth, which may be achieved by promoting the lateral root growth and inhibiting the primary root elongation. By testing the cell type-specific developmental markers, we demonstrated that Bacillus sp. LZR216 increases the DR5::GUS and DR5::GFP expression but decreases the CYCB1;1::GUS expression in Arabidopsis root tips. Further studies indicated that LZR216 is able to inhibit the meristematic length and decrease the cell division capability but has little effect on the quiescent center function of the root meristem. Subsequently, it was also shown that LZR216 has no significant effects on the primary root length of the pin2 and aux1-7 mutants. Furthermore, LZR216 down-regulates the levels of PIN1-GFP, PIN2-GFP, PIN3-GFP, and AUX1-YFP. In addition, the wild-type Arabidopsis seedlings in the present of 1 or 5 µM NPA (an auxin transport inhibitor) were insensitive to LZR216-inhibited primary root elongation. Collectively, LZR216 regulates the development of root system architecture depending on polar auxin transport. This study shows a new insight on the ability of beneficial endophytic bacteria in regulating postembryonic root development.