Screening of a microbial consortium with efficient corn stover degradation ability at low

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不同基质配比下黄瓜根际细菌群落结构及多样性分析

不同基质配比下黄瓜根际细菌群落结构及多样性分析

热带作物学报2024, 45(3): 641 652Chinese Journal of Tropical Crops不同基质配比下黄瓜根际细菌群落结构及多样性分析陈宏昊,唐小付*,聂圣贤,林佳佳,魏西,刘婷,任昊奎广西大学农学院,广西南宁 530004摘要:分析菇渣、锯末不同比例配比下黄瓜植株根际细菌群落结构特征发现,菇渣和锯末复配基质可替代常规商品基质泥炭(CK)栽培黄瓜,为新型基质栽培模式的应用提供依据。

与CK相比,用菇渣、锯末按照3种不同比例复配组成的基质栽培黄瓜,其产量、根际细菌多样性和丰富度指数间均显著提高,同时还影响根际细菌的代谢功能,其中菇渣与锯末比例为1∶1时(T2)表现最佳。

在门分类水平上,菇渣、锯末复配基质栽培条件下黄瓜植株根际中变形菌门(Proteobacteria)细菌丰度占比均低于CK,但拟杆菌门(Bacteroidota)细菌丰度占比均高于CK,且拟杆菌门(Bacteroidota)是菇渣、锯末复配基质栽培共有的优势细菌门类。

在属分类水平上,unclassified_c__Gammaproteo-bacteria、利姆诺杆菌属(Limnobacter)和朱氏杆菌属(Chujaibacter)是CK条件下黄瓜植株根系特有的优势细菌属;慢生根瘤菌属(Bradyrhizobium)、短波单胞菌属(Brevundimonas)、Dokdonella属是菇渣与锯末比例为1∶3(T1)处理下特有的优势细菌属;菇渣与锯末比例分别为1∶1(T2)和3∶1(T3)条件下黄瓜植株根际未发现特有的优势细菌属。

通过物种Venn分析,在种分类水平上,菇渣与锯末复配比例在1∶3(T1)、1∶1(T2)和3∶1(T3)处理下均增加了黄瓜植株根际细菌种分类水平数量,提升了根际细菌丰富度和多样性。

关键词:黄瓜;高通量测序;细菌群落结构;功能预测中图分类号:S642.2 文献标识码:AAnalysis of Cucumber Rhizosphere Bacterial Community Structure and Diversity under Different Matrix RatiosCHEN Honghao, TANG Xiaofu*, NIE Shengxian, LIN Jiajia, WEI Xi, LIU Ting, REN HaokuiCollege of Agronomy, Guangxi University, Nanning, Guangxi 530004, ChinaAbstract: The structural characteristics of the rhizosphere bacterial communities of cucumber plants under different proportions of mushroom residue and sawdust were analyzed, and it was found that mushroom residue and sawdust compound substrate could replace conventional commercial matrix peat (CK) cultivation cucumber, which providing a basis for the application of a new substrate cultivation mode. Compared with CK, cucumber cultivated with substrate composed of mushroom residue and sawdust in three different proportions significantly increased its yield, rhizosphere bacterial diversity and richness index, and also affected the metabolic function of rhizosphere bacteria, among which the ratio of mushroom residue to sawdust was 1∶1 (T2). At the classification level, the proportion of bacterial abundance in Proteobacteria of cucumber plants under the cultivation of mushroom residue and sawdust compound substrate was lower than that of CK, but the proportion of bacterial abundance of Bacteroidota was higher than that of CK, and Bac-teroidota was the dominant bacterial phylum shared by mushroom residue and sawdust compound substrate cultivation. At the taxonomic level, unclassified_c__Gammaproteobacteria, Limnobacter and Chujaibacter were the dominant bac-terial genera unique to the root system of cucumber plants under CK conditions. Bradyrhizobium, Brevundimonas, and Dokdonella were the dominant bacterial genera with a 1∶3 (T1) ratio of mushroom residue to sawdust. Under the con ditions of 1∶1 (T2) and 3∶1 (T3) ratios of mushroom residue to sawdust, respectively, no dominant bacterial genus收稿日期 2022-11-03;修回日期 2023-01-05基金项目 广西重点研发计划项目(桂科AB1850031,桂科AB19245016,桂科AB23026097)。

利用作物生长模型和时序信号甄别水稻镉胁迫

利用作物生长模型和时序信号甄别水稻镉胁迫

第37卷第4期农业工程学报 V ol.37 No.42021年2月Transactions of the Chinese Society of Agricultural Engineering Feb. 2021 249 利用作物生长模型和时序信号甄别水稻镉胁迫孔丽,刘美玲※,刘湘南,邹信裕(中国地质大学(北京)信息工程学院,北京 100083)摘要:在自然农田生态系统中,农作物的生长通常受到各类环境胁迫(如重金属胁迫、病虫害、水分、营养)的影响,如何区分重金属胁迫与其他胁迫有待进一步研究。

该研究选取了湖南省株洲为试验区,收集2017—2019年的Sentinel-2卫星影像数据,结合野外实测数据,开展水稻重金属镉(Cd)胁迫识别研究。

首先,利用作物生长模型World Food Studies (WOFOST)同化时序遥感数据获取每年的叶面积指数(Leaf Area Index,LAI)时间序列曲线;然后运用集合经验模态分解(Ensemble Empirical Mode Decomposition,EEMD)方法对LAI时间序列进行多尺度分解,得到不同的时序信号分量(Intrinsic Mode Function,IMF);最后使用动态时间规整(Dynamic Time Warping,DTW)方法计算受胁迫水稻分解后的时间序列与健康水稻分解后的时间序列之间的DTW距离,即归一化胁迫指数。

结果表明:归一化胁迫指数是水稻重金属胁迫敏感的参数,与土壤重金属含量的相关系数为0.851,水稻受到的胁迫程度越高,归一化胁迫指数值越大,反之越低;在试验区中,水稻重度重金属胁迫的分布面积比例相对较低,且主要集中在西部、东北部以及偏东南地区。

融合集合经验模态分解和动态时间规整方法能有效地甄别并定量分析水稻重金属胁迫状况,从而为作物重金属污染胁迫监测提供重要参考。

关键词:遥感;模型;重金属;镉胁迫;时序信号分解;WOFOSTdoi:10.11975/j.issn.1002-6819.2021.04.030中图分类号:S127 文献标志码:A 文章编号:1002-6819(2021)-04-0249-08孔丽,刘美玲,刘湘南,等. 利用作物生长模型和时序信号甄别水稻镉胁迫[J]. 农业工程学报,2021,37(4):249-256.doi:10.11975/j.issn.1002-6819.2021.04.030 Kong Li, Liu Meiling, Liu Xiangnan, et al. Identifying heavy metal (Cd) stress in rice using time-series signals and crop growth model[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2021, 37(4): 249-256. (in Chinese with English abstract) doi:10.11975/j.issn.1002-6819.2021.04.030 0 引 言随着经济的不断发展,工业化不断推进,土壤重金属污染成为当今世界面临的重大生态环境问题之一[1]。

牲畜粪便与麦秆混合厌氧发酵的产气量、发酵时间及最优温度

牲畜粪便与麦秆混合厌氧发酵的产气量、发酵时间及最优温度

牲畜粪便与麦秆混合厌氧发酵的产气量、 发酵时间及最优温度 !
张翠丽 # 李轶冰 # 卜东升 # 杨改河
!, " $, " % $, " !!
( ! 西北农林科技大学资源环境学院,陕西杨凌 &!$!’’ ;$ 西北农林科技大学农学院,陕西杨凌 &!$!’’ ;" 陕西省循环农业工 程技术研究中心,陕西杨凌 &!$!’’ ;% 新疆生产建设兵团农一师农业科学研究所,新疆阿拉尔 (%""’’ )
[ #* ]
" 因此, 单一以秸秆
作为沼气的发酵原料一直不被农民接受" 以粪便和 秸秆混合发酵可以有效地弥补单一秸秆作为发酵原 料的弊端, 同时还可以解决农村沼源短缺的问题, 但 目前有关粪便与秸秆配比发酵的系统研究还较为缺 乏" 温度是影响厌氧发酵的重要因素" 早在 +* 世纪 %* 年代, ,-./011 等 要" 吴满昌等
应 用 生 态 学 报# $’’( 年 ( 月# 第 !8 卷# 第 ( 期# # # # # # # # # # # # # # # # # # # # # # # # # # # # # ?K/FL.L \+@NF,- +R :MM-/L0 XO+-+GQ,:@G) $’’( , ;< (() : !(!&7!($$
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研究厌氧微生物与产气条件
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之间的关系时发现, 在厌氧发酵过程中温度尤为重 、 赵杰红等 的研究表明, 在温度对厌氧发酵降解程度的影响和温 度空间变异方面也进行了许多研究
[ #2 $ #( ]

纳米氧化铁与氧化剂对多环芳烃污染农田土壤修复和蔬菜健康风险的影响

纳米氧化铁与氧化剂对多环芳烃污染农田土壤修复和蔬菜健康风险的影响

纳米氧化铁与氧化剂对多环芳烃污染农田土壤修复和蔬菜健康风险的影响*第一作者:周佳靖,女,1996年生,硕士研究生,主要从事土壤修复研究。

通讯作者。

*山东省自然科学基金资助项目(No.ZR2020MD107、No.ZR2017MC068)。

周佳靖1柳修楚1郭 瑾1陈小宇1柴超1葛 蔚(1青岛农业大学资源与环境学院,山东 青岛266109 &.青岛农业大学生命科学学院,山东 青岛266109%摘要采用纳米氧化铁和氧化剂(过硫酸钠、H .O 2)联合技术修复多环芳烃(PAHs )污染农田土壤,分析纳米氧化铁与氧化剂联合修复对小白菜(Brassica chinensis L.)生长、PAHs 富集的影响,并进行健康风险评估#结果表明:(1)纳米氧化铁(2.0 g/kg )和H .O.C g/kg )联合修复对土壤、小白菜中PAHs 的去除效果最好,土壤中PAHs 去除率可达32.9%,小白菜地下部和地上部PAHs去除率分别为38.8%和38.9% # $)纳米氧化铁和过硫酸钠联合修复对小白菜生长存在抑制作用# $)经纳米氧化铁(2.0 g/kg )单独修复或纳米氧化铁$.0 g/kg )和H 2O 2$ g/kg )联合修复后,小白菜地上部中PAHs 对青少年和女性老年人的潜在致癌风险不再 存在#关键词纳米氧化铁过硫酸钠H .O .多环芳烃修复风险DOI :1015985/ki1001-3865.2021.02.016Effects of nano-Fe 2 O 3 and oxidants on soil remediation and health risk of polycyclic aromatic hydrocarbon in vegetablefrom contaminated farmland ZHOU Jiajing 1 LIU Xiuchu 1 ,GUO Jin 1 , CHEN Xiaoyu 1 , CHAI Chao' , GE Wei 2.(1.College of Resources and Environment Qingdao Agricultural University Qingdao Shandong 266109 ; ..College ofLif#Sci#nc#s "QingdaoAgriculturalUniv#rsity "QingdaoShandong 266109)Abstract : Remediationofpolycyclicaromatichydrocarbons (PAHs )contaminatedfarmlandsoilbynano-ferric oxidecombined withoxidants (sodiumpersulfate "H 2O 2 )wasstudied.Thee f ectsofnano-ferricoxidecombinedwith oxidantonthegrowthandPAHsaccumulationofpakchoi (Bra s icachinensis L .)wereanalyzed andthehealthriskwasassessed.Theresultsshowedthat :(1)thecombinedremediationofnano-ferricoxide (2.0g /kg )and H 2O 2 (2g/kg ) had the best effect on the removal of PAHs in soil and pakchoi. The removal rate of PAHs in soil was 32.9 % , and the reduction rates of PAHs in the underground and aboveground parts of pakchoi were 38. 8 % and 38. 9 % , respecively. (2) The combined remediaEion ofnano-ferric oxideand sodium persulfaEe inhibi edEhe growEh of pakchoi. (3) AfEernano-ferricoxide (2.0g /kg ) aloneornano-ferricoxide (2.0g /kg ) combinedwiEhH 2O 2 (2g /kg )"EherewasnoEhepoEenialcarcinogenicriskofPAHsinEheabovegroundparEsofpakchoiEoEeen-agersandfemaleseniors.Keywords : nano-ferric oxide ; sodium persulfate ; H2O2; polycyclic aromatic hydrocarbons ; remediation ; risk多环芳烃(PAHs )是环境中最普遍的有机污染物 之一,-,具有较高的遗传毒性与致癌性⑵。

盐碱地盐生植物根际耐盐促生菌的筛选及鉴定

盐碱地盐生植物根际耐盐促生菌的筛选及鉴定

农业科学研究Journal of Agricultural Sciences

第42卷第

4期Vol.42 No.42021年

12月

Dec. 2021

文章编号:1673-0747(2021)04-

0001-06

盐碱地盐生植物根际耐盐促生菌的筛选及鉴定梁翔宇,顾欣

刘文辉,

孙小涵,

白宇馨

马艳,李茜,赵莹(宁夏大学 农学院,

宁夏银川

750021)

摘 要:为开发利用盐生植物根际耐盐碱促生菌,选择宁夏银北地区盐碱地3种典型盐生植物:盐爪爪(Kalidiumfo- liaium)^碱蓬(Suaeda glauca)和芦苇(

Phragmites australis ),

利用培养法从其根际土壤中分离筛选耐盐威植物促生

菌。结果表明:从盐生植物根际土壤中分离获得可培养细菌83株,其中具有固氮功能的细菌23株,解磷菌

19株

,解

钾菌14株

,产IAA细菌15株,产铁载体细菌11株,具有ACC脱氨酶活性细菌10株。经过耐盐碱能力检测,

PJ10、

HL3和HY5菌株在(NaCl)=8%和pH=9的培养基中仍能生长。经分类鉴定,3株菌分别为壁芽葩杆菌(Poe讹iw

muralis)、短小芽葩杆菌(B pumilus)和萎缩芽葩杆菌(B airophaeus),其可作为新型微生物肥料的菌种。

关键词:盐生植物;盐緘地;根际促生菌中图分类号:Q939.96 文献标志码:A

盐碱化是全球耕地土壤面临的重要问题之一叫 为了利用盐碱化耕地并减少土壤盐碱化程度,人们

采用了许多土壤改良方法。其中,生物措施被认为是 较为有效且环境友好的措施叫例如,使用生物有机 肥可显著改善盐碱地的土壤性质叫施用菌剂可以 促进作物在盐碱化土壤中生长等I,生物有机肥和 菌剂中的耐盐碱有益功能菌发挥了重要作用叫 因此,分离获得耐盐碱性良好、可促进植物生长的功 能微生物对盐碱地改良及新型菌剂的开发具有重要 意义。根际促生菌(plant growth promoting rhizobacte- ria, PGPR)是指存在于植物根际周围的一类可以促 进植株生长,提高植株抗病性的微生物叫其对植物 生长和土壤改良均具有重要作用叫针对盐碱地植 物生长和土壤改良需求,耐盐碱的PGPR逐渐成为 研究热点。有研究者利用三级筛选体系从新疆碱蓬 (Suaeda glauca)根际土壤中分离获得PGPR菌,包 括芽抱杆菌(鸟“弘血sp.)、葡萄球菌(Staphylococcus sp.)、盐芽抱杆菌{Halobacillus sp.)、肠杆菌(EzUero- bacter sp.)和节杆菌(Arthrobacter sp.)回;从滨海盐碱 地的非盐生植物根际土壤中筛选获得了耐盐促生芽 抱杆菌 B. megaterium T1 -8 菌株和 B. pareflexus T4-9菌株㈣;从新疆盐爪爪(Kalidium foliatum )根际 土壤中分离获得了 57株细菌,分属4个门12个属,且 多数菌株兼有耐盐和促生作用问。以上研究说明,盐 碱地土壤中具有丰富的PGPR资源,有待进一步开发。银川平原位于我国西北部,由于气候、地质和人 为干扰等原因,土壤易发生盐碱化问。因此,当地对

橙色红曲菌乳清苷_5_磷酸脱羧酶基因的克隆

橙色红曲菌乳清苷_5_磷酸脱羧酶基因的克隆

-./01.2 ( -7=8<=8U8
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.D:Q7=UND<7E:;< 2L:9<L ) 2-5 结 合 2 !"# , 基 因 全
!=#" 实验方法
!"’"!( 简并引物的设计 ( 根据在 3->T 上已经公布
图 ." +K’F($F; !: O!. 的 +A3 产物; .: O!.?@ 的 +A3 产物; /: O!.?8 的 +A3 产物; 7: O!.@8 的 +A3 产物。!" 结果与分析
!#$" !"#% 基因片段的克隆与分析
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硫酸盐还原菌在顶空体积变化条件下所致镍的微生物腐蚀研究

硫酸盐还原菌在顶空体积变化条件下所致镍的微生物腐蚀研究

表面技术第53卷第4期硫酸盐还原菌在顶空体积变化条件下所致镍的微生物腐蚀研究田原,蒲亚男,孙天翔,侯苏,陈守刚*(中国海洋大学 材料科学与工程学院,山东 青岛 266100)摘要:目的通过改变厌氧瓶内顶空体积,研究硫酸盐还原菌(SRB)对金属镍的微生物腐蚀(MIC)机理,进而为镍基合金的腐蚀防护提供依据。

方法通过生物学检测技术、表面分析技术和电化学技术,评估了金属镍的MIC行为。

结果随着顶空体积的增大,更多的H2S以气体的形式析出到顶空,液相中硫化物浓度越低,SRB浮游和固着细胞数越高,点蚀坑越深,镍的腐蚀速率越高。

在200 mL的固定培养基体积下,顶空体积为90 mL和450 mL的镍试样失重分别是10 mL时的1.1倍和1.4倍,相应的点蚀坑深度分别增加了1.6倍、2.3倍。

在孵育7 d后,顶空体积为450 mL时低频阻抗模值最低,同时获得最大的腐蚀电流密度,达到7.64×10–6 A·cm–2。

结论利用细胞外镍氧化释放的电子在SRB细胞质中进行硫酸盐还原在热力学上是有利的,SRB所导致的镍的腐蚀属于EET-MIC。

关键词:镍;微生物膜;硫酸盐还原菌;顶空体积;微生物腐蚀;点蚀中图分类号:TG174 文献标志码:A 文章编号:1001-3660(2024)04-0068-09DOI:10.16490/ki.issn.1001-3660.2024.04.006Microbiologically Influenced Corrosion of Nickel Caused by Sulfate-reducing Bacteria under Different Levels of Headspace VolumeTIAN Yuan, PU Yanan, SUN Tianxiang, HOU Su, CHEN Shougang*(School of Materials Science and Engineering, Ocean University of China, Shandong Qingdao 266100, China)ABSTRACT: Nickel (Ni), an important alloying element, has been widely added to alloys to improve their corrosion resistance, and strengthened Ni-based alloys are commonly used in industries such as oil, gas, and offshore engineering, but still face the microbiologically influenced corrosion (MIC). The volume ratio of headspace/broth can affect microbial activity, thereby affecting the metabolism of biofilm and the process of extracellular electron transfer (EET). The MIC mechanism of Ni induced by sulfate-reducing bacteria (SRB) with varied headspace volume was investigated to provide a basis for the corrosion protection of Ni-based alloys. The MIC behavior of Ni was evaluated by biological detection, surface analysis, and electrochemical analysis techniques. Different headspace volume of 10 mL, 90 mL, and 450 mL was regulated with glass bottles and glass beads of different sizes with a fixed broth volume of 200 mL. The sessile cell count of Desulfovibrio vulgaris was 107 cells/cm2 after the 3 d pre-growth. The D. vulgaris sessile cell count on Ni coupons in the anaerobic bottles with 10 mL收稿日期:2022-12-22;修订日期:2023-03-30Received:2022-12-22;Revised:2023-03-30基金项目:国家自然科学基金项目(5197229);国防科技重点实验室基金项目(JS220406)Fund:National Natural Science Foundation of China (5197229); Key Laboratory of National Defense Science and Technology (JS220406)引文格式:田原, 蒲亚男, 孙天翔, 等. 硫酸盐还原菌在顶空体积变化条件下所致镍的微生物腐蚀研究[J]. 表面技术, 2024, 53(4): 68-76.TIAN Yuan, PU Yanan, SUN Tianxiang, et al. Microbiologically Influenced Corrosion of Nickel Caused by Sulfate-reducing Bacteria under Different Levels of Headspace Volume[J]. Surface Technology, 2024, 53(4): 68-76.*通信作者(Corresponding author)第53卷第4期田原,等:硫酸盐还原菌在顶空体积变化条件下所致镍的微生物腐蚀研究·69·headspace volume was 1.28×107 cell/cm2. The sessile cells count on Ni coupons in the 90 mL and 450 mL headspace volume increased by 213% and 288%, respectively compared with the 10 mL headspace. The fluorescence microscopy (FM) images also showed a relatively robust biofilm covering the Ni coupon surface. The corrosion rate of Ni coupon was analyzed by weight loss data and the maximum pitting depth distribution statistics (with a confocal laser scanning microscope, CLSM). The weight loss of Ni coupons with a headspace volume of 90 mL (1.1 mg/cm2) and 450 mL (1.4 mg/cm2) was 1.1 times and 1.4 times that of Ni coupons with the 10 mL headspace (1.0 mg/cm2), respectively. The maximum pit depth after the 7 d incubation was2.7 µm with 10 mL headspace volume, and the corresponding pitting depth increased 1.6 times (4.2 µm) and 2.3 times (6.1 µm)for 90 mL and 450 mL headspace, respectively. With a fixed broth volume of 200 mL in all bottles, a larger headspace meant that more H2S escaped to the headspace, which reduced the toxicity of H2S in broth to D. vulgaris. The larger the headspace volume, the higher the D. vulgaris sessile cells count, the deeper the pitting pits, and the higher the corrosion rate. The lowest polarization resistance (R p) value during the 7 d incubation period occurred in the headspace of 450 mL. Meanwhile, the lowest low-frequency resistance was obtained at a headspace volume of 450 mL. Corrosion current density (J corr) from potentiodynamic polarization data after the 7 d incubation with the 450 mL headspace (7.64× 10–6 A·cm–2) was more than 1.5 times and 2.1 times of that for headspace volume of 10 mL and 90 mL, respectively. The corrosion thermodynamic analysis and experimental data indicated that D. vulgaris utilized elemental Ni to replace the organic carbon source (lactate) as the electron donor. It is thermodynamically favorable to use the electrons released by extracellular Ni oxidation to reduce sulfate in D. vulgaris cytoplasm. Thus, Ni MIC by D. vulgaris belongs to EET-MIC.KEY WORDS: Ni; biofilm; sulfate-reducing bacteria; headspace; microbiologically influenced corrosion; pitting corrosion腐蚀是大多数物质与周围环境相互作用的电化学过程,每年因腐蚀问题会造成巨大的经济损失和安全问题。

玉米赤霉烯酮降解酶ZHD101的表达优化及在玉米浆脱毒中的应用

玉米赤霉烯酮降解酶ZHD101的表达优化及在玉米浆脱毒中的应用

第43卷第6期2020年11月河北农业大学学报JOURNAL OF HEBEI AGRICULTURAL UNIVERSITYVol.43 No.6Nov.2020玉米赤霉烯酮降解酶ZHD101的表达优化及在玉米浆脱毒中的应用陈 伟1,谷新晰1,李广靖1,徐 冉1,谷子林2,陈宝江2,卢海强1(1.河北农业大学 食品科技学院,河北 保定 071000;2. 河北农业大学 动物科技学院,河北 保定 071000)摘要:本研究旨在提高玉米赤霉烯酮降解酶ZHD101表达效率及探讨其在玉米浆脱毒中的应用潜力。

ZHD101基因在优化后共27个稀有密码子被同义替换,GC平均值由56.35%调整为60.23%,CAI值由0.64调整为0.97,优化前后基因序列的一致性为81.1%,重组酶ZHD101成功高效表达。

随后采用Plackett-Burman设计和响应面分析法明确工程菌在种龄OD值为1.0,诱导温度30.0℃和诱导8.5 h条件下,酶的表达量最高,提高了约3倍左右。

玉米赤霉烯酮降解酶ZHD101对玉米浆的脱毒率随反应时间延长而提高,在反应1、2和3 h对玉米浆中玉米赤霉烯酮的降解率分别为(6.2±1.5)%、(13.4±2.8)%和(17.1±8.0)%。

本研究可为玉米赤霉烯酮降解酶ZHD101的生产和玉米赤霉烯酮毒素脱毒的实际应用提供一定的科学指导。

关 键 词:玉米赤霉烯酮;高效表达;生物降解;玉米浆;食品安全中图分类号:Q814.4 开放科学(资源服务)标识码(OSID):文献标志码:AOptimized expression of a zearalenone degrading enzyme (ZHD101) and the application in detoxification of corn steep liquor CHEN Wei1, GU Xinxi1, LI Guangjing1, XU Ran1, GU Zilin2, CHEN Baojiang2, LU Haiqiang1(1.College of Food Science and Technology, Hebei Agricultural University, Baoding 071000, China;2. College of Animal Science and Technology, Hebei Agricultural University, Baoding 071000, China)Abstract: The aim of this study is to improve the substantial application and expression efficiency of zenrlenoredegrading enzyme(ZHD101). After optimization, 27 rare codons of ZHD101 gene were replaced synonymously,the average GC value was adjusted from 56.35% to 60.23%, the CAI value was adjusted from 0.64 to 0.97, theconsistency of gene sequence before and after optimization was 81.1%, and the recombinant enzyme ZHD101was successfully and efficiently expressed. Subsequently, Plackett-Burman design and response surface analysiswere used to determine that the expression of the enzyme was the highest under the conditions of OD value of 1.0,induction temperature of 30.0 C and induction time of 8.5 h, which increased about 3 times. The detoxification rateof zearalenone-detoxifying lactonase ZHD101 to corn syrup increased with the extension of reaction time. Thedegradation rates of zearalenone in corn steep liquor were (6.2±1.5)%, (13.4±2.8)% and (17.1±8.0)% respectively收稿日期:2020-04-22基金项目:河北省高等学校科学技术研究项目(QN2018005);河北省重点研发计划项目(20236613D).第一作者: 陈伟(1994-),湖北黄冈人,男,硕士研究生,主要从事食品微生物资源的开发与应用.E-mail:****************通信作者: 卢海强(1981-),河北保定人,男,博士,副教授,从事食品酶工程的教学与科研工作.E-mail:********************.cn本刊网址:http: // hauxb. hebau. edu. cn: 8080 /CN/ volumn / home. shtml文章编号:1000-1573(2020)06-0026-09DOI:10.13320/ki.jauh.2020.010927第6期in 1, 2 and 3 hours of reaction time. This study provides the scientific guidance for the production of zearalenone degrading enzyme ZHD101 and the practical application of zearalenone toxin detoxification.Keywords: zearalenone; high-level expression; biodetoxifification; corn steep liquor; food safety玉米浆是湿法工艺生产玉米淀粉的主要副产品,是1种黏稠酸性的浆体[1]。

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Journal of Integrative Agriculture 2016,15(10):2369-2379 Available online at www.sciencedirect.com Sc enceDirect l 

RESEARCH ARTICLE ^一一一一一! ‘ 一一■一一一■一i■ ●i i一 ,,一i Creenlng o1'a mlCroDial conSortIUm with e竹icient corn stover degradation ability at low temperature 

Qinggeer’,GAO Ju—lin’,YU Xiao-fang’,ZHANG Bao-Iin ,WANG Zhi.gang’,Bodigin Naoganchaolu’, 

HU Shu-ping’,SUN Ji-ying’,XIE Min’

,WANG Zhen’ 

Agricultural College,Inner Mongolia Agricultural University,Hohhot 010019,P.R.China Chemistry and Environmental Science College,Inner Mongolia Normal University,Hohhot 010019,P.R.China 

Abstract To speed up the degradation of corn stover directly returned to soil at Iow temperature,the corn stOver_degradinq micr bial consortium GF-2O,acclimated to biological decomposition in the frigid region.was successfully constructed under long-term limiting substrate.To evaluate its potential in accelerating the decomposition of un—pretreated corn stover.t} decomposing property.fermentation dynamic and the microbial diversity were analyzed.GF一20 degraded corn stover I 32%af【er 1 5-day fermentation at 1 O。C.Peak activities of filter paperlyase(FPA),p-glucosidases(CB),endoglucanas ̄ (Cx),and ceIlobi0hydr0Iases(C1)were 1.15,1.67,1-73,and 1.42 U mL一’,appearing atthe 6th,3rd,11th,and 9th d,respe tively.The pH averaged at 6.73_8.42,and the optical density(OD)value peaked at 1.87 at the 1 20 h of the degradatk process.Cellulase,hemicellulase and lignin in corn stover were persistently degraded by 44.85.43.85 and 25.29% the end of incubation.Result of denaturing gradient gel electrophoresis(DGGE)profiles demonstrated that GF-20 had stable component structure under switching the temperature and pH.The composition of the GF一20 was also analyze by constructing bacteriaI 1 6S rDNA clone Iibrary and fungal 1 8SrDNA-PCR—DGGE.Twenty-two bacteriaI clones and fo fungaI bands were detected and identified dominant bacteria represented by Cellvibrio m fus subsp..Azospira oryza Arcobacter defluyii.and CIostridium populefi and the fungi were mainly identified as related to osporon sp. 

Keywords:corn stover,degradation,microbial consortium,low temperature 

1.1ntroduction cellulosic biomass has been source for renewable energy recently recognized as a (Sukumaran et a1.2009; 

Received 24 July。201 5 Accepted 1 1 January,201 6 COrresp0ndence GAO Ju-lin,Tel:+86—471—4301317, E—mail:nmga ̄ulin@1 63.com 

@201 6.CAAS.All rights reserved.Published by Elsevier Ltd doi:10.1016/¥2095-31 19(15)61272-2 

Honeina et a1.20 1 21.Transformation of corn stover to n trient substance considerably reduces waste accumulatk in the environment,thereby serving as a means to produ( value.added products.In China.the totaI annuaI productk of crop straw was estimated to be 700 million t(Bi et{ 201 1 1,221.56 million t of which was corn stover.Howew only a small portion of stovers were used to produce anim feed or organic fertilizer,most stovers were burnt(War ef a1.201 1).Corn stover returned on field(38.99 million accounts for only 1 7.6%ofthe corn residues fL(】ef a1.201: The northeast region.where the harvest is 54-79 millior of corn.with 68.49 million t of stover,accounting for 31 of the totaIin the country,is the largest corn belt in C:hin 2370 Qinggeereta1./JournalofIntegrativeAgriculture 2016,15(10):2369-2379 However.Iess than 1 0%stover returned on field because of lower temperature,drought windy and dusty weather (L(】ef a1.201 3),and unsatisfactory decomposition impacts planting and seedling emergence in the following year,and aggravates plant diseases and insect pests. Therefore.acceleration of stover biologicaI decay in the field is badly in needed in northeastern China.Many microorganisms which are characterized by multicompo— nent,cellulolytic enzyme systems have been reported with cellulosic activities.Cellulolytic bacteriaI species include Fmrobacters Actinomycetes.Cellulomonas Cellvibrio. Cytophaga,Bacillus,Paenibacillus sp.,Clostridium thermo— ceflum.Caldicellulosirup幻rohsidians ̄.Methanobrevibacter ruminanfium.etc.tFreier et a1.1988;Kirclman et a1.2002; Zhanfl et a1.2008;Lc】ef a1.2011:Maki et a1.2011:Miran— da et a1.2011:Wu and He 2015),fungaI species include Aspergillus niger.Trichoderma reesei,Trichoderma viride. Penicillium.Trichoderma sp..etc.(Li et a1.201 3:0keke ef a1.2014:Strakowska et a1.2014:Guer『aIi ef a1.201 5). However.jt has been reported that the mixture microbiaI composite showed rather higher effective decomposition capacity and cellulase activity compared with any single strains(Haruta et a1.2002;Guo et a1.2008).But previous investiaatiOns on microbial communities were conducted at medium—high temperature fGuo et a1.2008;Li et a1. 201 1),not appropriate for low temperature.There are still no reports on microbiaI communities which can efficiently hydrolyze un—pretreated naturaI IianOcellulases at Iow tern— perature.In order to accelerate the decomposition of corn stover,a mixture of microbes,which had a higher enzyme activity at low temperature,was specifically grown and selected in a limiting substrate of corn stover. 2.Materials and methods 2.1.Experimental materials noculums sources of soil and cow dung were collected Table 1 Sample collecting sites and characteristics frOm 31 diferent sites in the cold regions of Northem China (Table 1).Each sample was placed in a sterile plastic bag, sealed and transported to the laboratory on ice and then stored in the dark at 4。C.Post—harvest corn stover was obtained from the experimental farm of Inner Mongolia Ag— riculturaI University.The stover,without any diseases and contaminating insects.was air-dried,washed with water, and then oven—dried at 80。C.For submerged fermentation, dried stover was cut into scraps approximately 1 cm in length and mixed thoroughly with the medium.The filter paper (10 cmx1 cm in length and width,about 0.1 g)used in this study was Whatman No.1 fJapan). The culture media were consisted of enrichment medium No.1 peptone cellulase solution(PCS)medium(Lfi et a1. 201 2),the corn stover as the carbon source(2%w/v),en— richment medium No.2(2 g(NH4)2SO4,1 g K2HPO4,0.5 g MgSO4"7H,O,2 g CaCO ,2 g NaCI,filter paper,and 1 000 mL distilled water(pH=7.5+0.1)).The basaI medium used in this study was the same as en richment medium No.2.All media were sterilized at 1 21。C f0r 20 min. 

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