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1. 光照强度对橘小实蝇性行为的调控作用
REN Cong, ZHANG Jie, YUAN Jin-xi, WU Yun-qi-qi-ge, YAN Shan-chun, LIU Wei, WANG Gui-rong
Journal of Integrative Agriculture    2023, 22 (9): 2772-2782.   DOI: 10.1016/j.jia.2023.04.025
摘要193)      PDF    收藏

橘小实蝇(Bactrocera dorsalis (Hendel))是柑橘类水果的毁灭性虫害。雌虫在成功交配后将卵产入成熟果实,导致其发霉腐烂失去经济价值,从而严重危害柑橘产业。自然条件下的橘小实蝇交配时间发生于黄昏时段,此时下降的光照强度是诱发其交配的关键条件。本研究首先通过设置0-30000lux共10种光照强度,从而确定何种光强能够明显调控橘小实蝇的交配行为。进一步选择了三种明显调控其行为的光照强度,测试了这些光照强度对雄虫求偶(振翅)及雌虫对性信息素2,3,5-三甲基吡嗪(2,3,5-trimethylpyrazine TMP)趋向性的影响。最后,在实验室中将强光和黑暗条件人工组合,测试其是否可以阻止橘小实蝇的交配,以期待为未来橘小实蝇的行为调控提供理论基础。结果表明,橘小实蝇成虫能在较低光照(<1000lux)正常交配,光强越强其交配数量越低,在光强达到20000lux以上时几乎无交配。较强光强明显减弱并推迟了雄虫的振翅行为与雌虫对TMP的趋向行为,不同的是雄虫在10000lux下仍有一定程度的振翅,而雌虫在此光强下对TMP几乎无趋向行为。成虫在无光情况下无交配行为,在此过程中雄虫无振翅行为而雌虫则对TMP失去趋向行为。进一步模拟不利光照条件,在强光10000lux一小时后持续无光,橘小实蝇成虫不进行交配。因此,光照条件是对橘小实蝇求偶交配的重要条件,未来可通过人工改变光强或其他手段干扰橘小实蝇成虫感弱光的分子靶标调控其求偶交配行为,从而开发新型绿色的橘小实蝇防控技术。

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2. JIA-2022-0120 具有优异面团品质特性的小麦-粘果山羊草1Uk(1A)代换系的创制及鉴定
JIANG Yun, WANG De-li, HAO Ming, ZHANG Jie, LIU Deng-cai
Journal of Integrative Agriculture    2023, 22 (4): 999-1008.   DOI: 10.1016/j.jia.2022.08.020
摘要239)      PDF    收藏

从野生近缘种中发掘新的高分子量谷蛋白亚基HMW-GS是改良小麦加工品质的有效途径。本研究旨在鉴定一份小麦-粘果山羊草渐渗系N124的染色体组成,并评价其对小麦品质相关性状的影响。荧光原位杂交FISH)核型分析表明,N124是一个1Uk(1A)二体代换系。十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和反相高效液相色谱(RP-HPLC发现N124表达了两个源自粘果山羊草的HMW-GSPacBio RNA测序和系统发育分析证实了这两个HMW-GS分别为UkxUky。与小麦亲本相比,N124除穗粒数较少外,无明显农艺性状缺陷,同时多项主要品质指标均有提高,表明N124可作为小麦品质改良的桥梁材料加以利用。

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3. JIA-2021-0045 QoIs和SDHIs类杀菌剂对禾谷镰刀菌线粒体动态平衡的影响依赖于FgDnm1
KANG Jin-bo, ZHANG Jie, LIU Yin-kai, SONG Ji-chang, OU Jian-lin, TAO Xian, ZHOU Ming-guo, DUAN Ya-bing
Journal of Integrative Agriculture    2023, 22 (2): 481-494.   DOI: 10.1016/j.jia.2022.08.118
摘要436)      PDF    收藏

禾谷镰刀菌(Fusarium graminearum)引起的小麦赤霉病(Fusarium head blight, FHB)是一种严重为害粮食作物的真菌病害,不仅引起作物产量损失与品质下降,而且病原菌在感病的谷粒中分泌的真菌毒素,严重威胁粮食安全。前期研究表明甲氧基丙烯酸酯类(quinone outside inhibitors, QoIs)和琥珀酸脱氢酶抑制剂类succinate dehydrogenase inhibitors, SDHIs)杀菌剂可破坏线粒体动态平衡,引起线粒体碎片化。动力蛋白和动力相关蛋白(DRPs)作为GTPase超家族成员,参与调控真核细胞线粒体分裂、囊泡出芽分裂等功能,但其在禾谷镰刀菌中的功能尚不清楚。在本研究中,我们利用BLAST分析发现禾谷镰孢菌中与酵母Dnm1的同源蛋白FgDnm1,并对其进行了生物学功能研究。结果表明,FgDnm1参与调控着菌丝生长、有性生殖及杀菌剂药敏性。此外,我们利用荧光标记技术和激光共聚焦显微镜发现FgDnm1与线粒体共定位,且参与调控禾谷镰刀菌产毒小体的结构形成及脱氧雪腐镰刀菌烯醇(DON)的生物合成。进一步研究表明,甲氧基丙烯酸酯类杀菌剂(QoIs)和琥珀酸脱氢酶抑制剂类杀菌剂(SDHIs)均会引起线粒体的碎片化,FgDnm1的缺失会导致线粒体呈现丝状网络分布,并阻断了QoIs和SDHIs诱导的线粒体碎片化。本研究揭示了线粒体动态平衡对禾谷镰刀菌菌丝生长发育、杀菌剂敏感性和毒素形成的影响。因此,我们推论QoIs与SDHIs杀菌剂引起的线粒体动态平衡变化依赖于FgDnm1。

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4. 一个控制油菜半矮杆性状的主效新位点BnaSD.C3的精细定位及候选基因分析
WANG Xiao-dong, CAI Ying, PANG Cheng-ke, ZHAO Xiao-zhen, SHI Rui, LIU Hong-fang, CHEN Feng, ZHANG Wei, FU San-xiong, HU Mao-long, HUA Wei, ZHENG Ming, ZHANG Jie-fu
Journal of Integrative Agriculture    2023, 22 (10): 2981-2992.   DOI: 10.1016/j.jia.2023.02.017
摘要217)      PDF    收藏

株高是影响甘蓝型油菜产量、收获指数和抗倒伏性的关键株型特征,然而,油菜株高的遗传调控机制仍不清楚。本研究利用EMS诱变获得了一个半矮杆突变体df34遗传分析结果表明,df34半矮杆性状由一半显性基因控制。利用BSA-Seq方法将目的基因定位到C3染色体上命名为BnaSD.C3。随后,利用图位克隆的方法,将BnaSD.C3精细定位到“Darmor-bzh基因组的297.35 kb区间内。然而,在“Darmor-bzh基因组上的这一区间内,没有潜在的调控株高性状候选基因结合基因组重测序、转录组测序、植物激素分析、结构变异分析和基因功能注释等信息,在“ZS11”参考基因组上,发现BnaC03G0466900ZSBnaC03G0478900ZSBnaSD.C3的重要候选基因。研究为甘蓝型油菜矮化及株型育种提供了新的基因资源,为解析甘蓝型油菜株高的遗传调控机制提供了新的见解。

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5. The transcription factor FgNsf1 regulates fungal development, virulence and stress responses in Fusarium graminearum
SHI Dong-ya, REN Wei-chao, WANG Jin, ZHANG Jie, Jane Ifunanya MBADIANYA, MAO Xue-wei, CHEN Chang-jun
Journal of Integrative Agriculture    2021, 20 (8): 2156-2169.   DOI: 10.1016/S2095-3119(20)63339-1
摘要154)      PDF    收藏

Nsf1(Nutrient and stress factor 1)是典型的C2H2型锌指蛋白,酿酒酵母中Nsf1在非发酵碳源或者盐胁迫的条件下才会表达。进化树分析发现该基因在不同物种间比较保守,然而,Nsf1的功能在丝状真菌中研究得并不是很透彻。为了探索FgNsf1在小麦赤霉病的病原菌禾谷镰孢菌中的功能,我们构建了FgNSF1基因敲除体(ΔFgNsf1)和包含GFP标记的回复体(ΔFgNsf1-C),亚细胞定位表明FgNsf1蛋白集中于细胞核。进一步研究发现,与野生菌株PH-1和回复体ΔFgNsf1-C 相比,敲除体ΔFgNsf1的菌丝体生长速率明显减慢,分生孢子产量及萌发率显著下降,且有畸形孢子产生,子囊壳产量也显著降低。但是红色镰刀菌素和黄色镰刀菌素的产量明显增加,为了验证这一结果,我们利用实时荧光定量PCR技术检测了相关基因(AurJAurFAurOAurR2)的表达量,研究结果发现,相关基因的表达量都显著上调。此外,使用不同浓度的NaCl处理时,野生菌株PH-1中FgNSF1基因的表达量均上调,而在使用不可发酵碳源乙醇、甘油或醋酸盐作为唯一碳源时,FgNSF1基因的表达量都显著下调。另外我们发现敲除体对渗透,细胞壁,氧化和部分金属离子等胁迫因子的敏感性显著下降,只对0.2M镁离子胁迫的敏感性显著提高。药敏性实验发现,敲除体对咯菌腈和抑菌脲的抗药性明显增强,而对戊唑醇和多菌灵的敏感性显著提高。随后,我们在小麦胚芽鞘和麦穗上进行了致病力实验,结果发现ΔFgNsf1的致病力显著减弱,在产毒培养基中DON(脱氧雪腐镰刀烯醇)产量也显著下降,以及DON毒素合成相关基因TRI5和TRI6的基因表达量也显著下调。结论:FgNSF1在禾谷镰孢菌的生长发育,有性和无性生殖,应对外界胁迫,产毒和致病的过程中扮演着重要的角色。创新性:我们首次系统地报道了FgNSF1在禾谷镰孢菌中的功能。


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6. Genetic parameter estimation and genome-wide association study (GWAS) of red blood cell count at three stages in a Duroc×Erhualian pig population
NAN Jiu-hong, YIN Li-lin, TANG Zhen-shuang, CHEN Jian-hai, ZHANG Jie, WANG Hai-yan, DU Xiao-yong, LIU Xiang-dong
Journal of Integrative Agriculture    2020, 19 (3): 793-799.   DOI: 10.1016/S2095-3119(19)62773-5
摘要135)      PDF    收藏
Red blood cells play an essential role in the immune system.  Moreover, red blood cell count (RBC) is an important clinical indicator of various diseases, including anemia, type 2 diabetes and the metabolic syndrome.  Thus, it is necessary to reveal the genetic mechanism of RBC for animal disease resistance breeding.  However, quite a few studies had focused on porcine RBC, especially at different stages.  Thus, studies on porcine RBC at different stages are needed for disease resistant breeding.  In this study, the porcine RBC of 20-, 33-, and 80-day old were measured, and genetic parameter estimation and genome-wide association study (GWAS) were both performed.  As a result, the heritability was about 0.6 at the early stages, much higher than that at 80 days.  Nine novel genome wide significant single nucleotide polymorphisms (SNPs), located at Sus scrofa chromosome (SSC)3, 4, 8, 9, 10 and 15, respectively, were identified.  Further, TGFβ2, TMCC2 and PPP1R15B genes were identified as important candidate genes of porcine red blood cell count.  So different SNPs and candidate genes were found significantly associated with porcine RBC at different stages, suggesting that different genes might play key roles on porcine RBC at different stages.  Overall, new evidences were offered in this study for the genetic bases of animal RBC, and that the SNPs and candidate genes would be useful for disease resistant breeding of pig.
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7.
One-time fertilization at first flowering improves lint yield and dry matter partitioning in late planted short-season cotton
LUO Hong-hai, WANG Qiang, ZHANG Jie-kun, WANG Lei-shan, LI Ya-bing, YANG Guo-zheng
Journal of Integrative Agriculture    2020, 19 (2): 509-517.   DOI: 10.1016/S2095-3119(19)62623-7
摘要132)      PDF    收藏
Cotton producers have substantially reduced their inputs (labor, nutrients, and management) mainly by adopting a short-season cropping management that is characterized by late sowing, high density, and reduced fertilization with one-time application at the first bloom stage without lint yield reduction.  However, it has been hypothesized that one-time fertilization at an earlier growth stage could be a more effective and economic management practice.  A two-year field experiment was conducted by applying five fertilizer one-time fertilization at 0 (FT1), 5 (FT2), 10 (FT3), 15 (FT4), and 20 (FT5) days after the first flower appeared in the field and one three-split fertilizer application taken as the conventional control (FT6), making six treatments altogether.  Cotton growth period, biomass accumulation, yield, and its formation were quantified.  The results showed that the one-time fertilization did not affect the cotton growth progress as compared to FT6, however, the total crop cycles for FT3–FT5 were 3 days shorter.  FT1 produced the highest cotton lint yield (1 396 kg ha–1), which was similar to the FT6 but higher than the other treatments, and could be attributed to more bolls per unit area and higher lint percentage. Cotton yield was positively correlated with cotton plant biomass accumulated.  FT1 had both the highest average (VT) (193.7 kg ha–1 d–1) and the highest maximum (VM) (220.9 kg ha–1 d–1) rates during the fast biomass accumulation period.  These results suggest that one-time fertilizer application at the first flower stage might be an adjustment that is more effective than at first bloom, and allowed for easier decision making for application date due to non counting of plants with flowers is needed.
 
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8. Alanine-substituted mutant on Gly373 and Asn375 of Cry1Ai-h-loop 2 causes reduction in both toxicity and binding against Helicoverpa armigera
LIU Yu-xiao, ZHOU Zi-shan, LIANG Ge-mei, SONG Fu-ping, ZHANG Jie
Journal of Integrative Agriculture    2019, 18 (5): 1064-1071.   DOI: 10.1016/S2095-3119(18)61933-1
摘要165)      PDF    收藏
Cry1Ai-h-loop 2 is a mutant of Cry1Ai constructed by exchanging loop 2 from Cry1Ah protein and shows insecticidal activity against Helicoverpa armigera.  The toxicity of Cry1Ai-h-loop 2, in contrast to the very low toxicity of Cry1Ai, is closely associated with the eleven residues in the loop 2 region.  To characterize the key sites of loop 2 in Cry1Ai-h-loop 2, alanine-substituted mutants were generated.  The toxicity of these mutants against H. armigera indicated that dual-mutant on Gly373 and Asn375 caused a significant decrease in toxic activity.  ELISA binding and competition binding assays demonstrated that the reduction of toxicity in the mutant of interest was correlated with decreased binding affinity.
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9. Transcriptional profile of gene clusters involved in the methylerythritol phosphate pathway in Bacillus subtilis 916
XIAO Ya-jing, GAO Tan-tan, PENG Qi, ZHANG Jie, SUN Dong-mei, SONG Fu-ping
Journal of Integrative Agriculture    2019, 18 (3): 644-655.   DOI: 10.1016/S2095-3119(18)62001-5
摘要206)      PDF(pc) (680KB)(162)    收藏
The methylerythritol phosphate pathway is responsible for the biosynthesis of terpenoids, the largest class of secondary metabolites.  Although the structures and functions of the proteins involved in this pathway have been well studied in Bacillus subtilis, only a few studies have reported the transcriptional profile of the genes involved.  Therefore, we analyzed methylerythritol phosphate pathway genes in the genome of B. subtilis 916, which has been developed as a biological control agent against some rice diseases in China.  Our results showed that methylerythritol phosphate pathway genes were distributed throughout the genome of this strain.  These genes were transcribed during both the exponential and stationary phases.  We further confirmed the transcription units of dxs, dxr, ispD, ispF, ipK, ispG, ispH, idi, and ispA in B. subtilis 916 through reverse transcription-PCR analyses; the results showed that these nine genes were located in seven different operons.  The transcript start sites of the seven different operons were determined by 5´-rapid amplification of cDNA ends-PCR.  Thus, our study provides a molecular basis at the transcriptional level for investigating homoterpene synthesis in the methylerythritol phosphate pathway of B. subtilis 916.
 
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10. Inheritance and molecular characterization of resistance to AHAS-inhibiting herbicides in rapeseed
HU Mao-long, PU Hui-ming, GAO Jian-qin, LONG Wei-hua, CHEN Feng, ZHOU Xiao-ying, ZHANG Wei, PENG Qi, CHEN Song, ZHANG Jie-fu
Journal of Integrative Agriculture    2017, 16 (11): 2421-2433.   DOI: 10.1016/S2095-3119(17)61659-9
摘要791)      PDF    收藏
Rapeseed is a very important oil crop in China; however, its production is challenging due to the absence of effective weed management strategies.  This is predominantly because of a shortage of herbicide resistance genes.  Acetohydroxyacid synthase (AHAS) herbicides inhibit AHAS, a key enzyme involved in branched-chain amino acid synthesis that is required for plant growth.  A rapeseed line designated M342 with AHAS herbicide resistance was developed through seed mutagenesis and was studied to assess the level and mode of inheritance of the resistance and to identify the molecular basis of resistance.  M342 possessed a high level of cross-resistance to sulfonylureas (SUs) and imidazolinones (IMIs).  This resistance was due to AHAS insensitivity to these herbicides and was inherited as a dominant trait conferred by a single nuclear-encoded gene.  Molecular analysis revealed the presence of a Trp574Leu mutation in M342, and an allele-specific cleaved amplified polymorphic sequence (AS-CAPS) marker was developed and cosegregated with herbicide resistance in the F2, BC1, and BC2 populations.  This mutation altered the transcript levels of BnAHAS1 and BnAHAS3 in M342 compared with those in the wild type, but it did not affect the agronomic or quality traits.  The simple genetic inheritance of this mutation and the availability of the cleaved amplified polymorphic sequence (CAPS) marker and herbicide resistance gene should facilitate the development of herbicide-resistant rapeseed cultivars for effective weed control in China.  
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11. Complete genome sequence of Bacillus thuringiensis Bt185, a potential soil insect biocontrol agent
LI Yan-qiu, SHU Chang-long, SHAN Yue-ming, GENG Li-li, SONG Fu-ping, ZHANG Jie
Journal of Integrative Agriculture    2017, 16 (03): 749-751.   DOI: 10.1016/S2095-3119(16)61422-3
摘要953)      PDF    收藏
Bacillus thuringiensis Bt185 and its insecticidal spectrum-expanded engineering strains are considered as potential biocontrol agents to soil insect Holotrichia parallela, Holotrichia oblita or Anomala corpulenta.  Here we reported the complete genome of strain Bt185, it harbors eight plasmids, and plasmid pBT1850294 carries three cry8 genes.
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12. Identification of similar transcriptional regulatory mechanisms in multiple cry genes in Bacillus thuringiensis HD12
SONG Zhi-ru, PENG Qi, SHU Chang-long, ZHANG Jie, SUN Dong-mei, SONG Fu-ping
Journal of Integrative Agriculture    2017, 16 (01): 135-143.   DOI: 10.1016/S2095-3119(16)61398-9
摘要690)      收藏
Bacillus thuringiensis subspecies morrisoni strain HD12, whose genome harbors multiple insecticidal protein-encoding genes, includes eight cry genes, as indicated by genome sequencing.  This strain produces crystals that are toxic to lepidopteran species.  These crystal inclusions were purified by sucrose gradients and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by liquid chromatography-mass spectrometry, and found to contain five proteins (Cry1Da, Cry1Ae, Cry1Bb, Cry1Fb, and Cry1Ja).  The transcriptional activities of the cry1Da, cry1Ae, cry1Bb, cry1Fb, and cry1Ja promoters indicated that transcription of cry1Da is controlled by SigE; however, the other four cry genes were found to be controlled by both SigE and SigK.  The activities of the cry1Ja and cry1Fb promoters were the strongest among the five genes studied.  These promoters were therefore used to direct the expression of the Cry1Ac- and Cry2Ab-encoding genes concurrently in a single strain.  Our findings indicate that promoters with the same transcriptional profile may be used to direct the expression of different cry genes in one strain.  Our results are expected to be valuable for the construction of strains with efficient expression of multiple cry genes in order to overcome current limitations associated with the application of B. thuringiensis-based insecticides.
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13. Separation and purification of deoxynivalenol (DON) mycotoxin from wheat culture using a simple two-step silica gel column chromatography
ZHAO Xiu-mei, LI Rong-jia, ZHOU Chuang, ZHANG Jie, HE Cheng-hua, ZHENG Ya-ting, WU Wen-da
Journal of Integrative Agriculture    2016, 15 (3): 694-701.   DOI: 10.1016/S2095-3119(15)61098-X
摘要1871)      PDF    收藏
Deoxynivalenol (DON) is a type B trichothecenes mycotoxin produced by several Fusarium species, often found in foodstuffs for humans and animals. DON is in great demand for the toxicological researches both in vivo and in vitro. In this work, wheat culture was inoculated with a Fusarium graminearum PH-1 strain for DON production. The solvent system for crude extraction was acetonitrile-water (84:16, v/v). A simple two-step silica gel column chromatography was employed to separate the DON mycotoxin from wheat culture, combined with preparative high performance liquid chromatography (preparative HPLC) to purify the compound. The solvent system for the second silica gel column chromatography was methylene chloride-methanol (17:1, v/v), which provided a good elution effect selected on thin layer chromatography (TLC). The target compound was identified by HPLC, and the chemical structure was confirmed by mass spectrometry (MS) and 1H and 13C nuclear magnetic resonance (NMR) spectroscopy. A total of 433 mg of purified DON was obtained from 1 kg of wheat culture, with a purity of 99.01%. The study had provided an easy-operating and cost-effective method to isolate an expensive compound in a simple way.
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14. Comparison and optimization of the method for Cry1Ac protoxin preparation in HD73 strain
ZHOU Zi-shan, YANG Su-juan, SHU Chang-long, SONG Fu-ping, ZHOU Xue-ping, ZHANG Jie
Journal of Integrative Agriculture    2015, 14 (8): 1598-1603.   DOI: 10.1016/S2095-3119(14)60950-3
摘要2095)      PDF    收藏
Bacillus thuringiensis is one of the most widely used bioinsecticides, and cry gene is the major insecticidal gene. Because Cry1Ac protein shows strong toxicity against many lepidopteran species, it has been applied widely in spraying products and transgenic Bt-crops. The preparation of Cry protoxin is the first step in the very important processes of understanding the insecticidal mechanism, resistance screening, and biosafety assessments. The media for crystal production and the method for Cry protoxin preparation were varied, however, it was not clear which was better for preparing a larger amount of Cry protoxin. In this paper, three media for crystal production and the method for Cry1Ac protoxin preparation from HD73 strain were compared to find an efficacious way to prepare a large number of Cry1Ac protoxin. The results showed that the 1/2 LB (Luria-Bertani) medium was the ideal medium for crystal production, because the total yield of Cry1Ac protoxin in 300 mL 1/2 LB medium was (112.38±5.64) mg, the highest one among three media; the repeated crystal solubilization method was better for the preparation of the Cry protoxin comparing with the continuous crystal solubilization method. It will be a reference for other Cry protoxin preparation, especially for larger number.
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15. Developing transgenic maize (Zea mays L.) with insect resistance and glyphosate tolerance by fusion gene transformation
SUN He, LANG Zhi-hong, LU Wei, ZHANG Jie, HE Kang-lai , ZHU Li, LIN Min, HUANG Da-fang
Journal of Integrative Agriculture    2015, 14 (2): 305-313.   DOI: 10.1016/S2095-3119(14)60855-8
摘要1803)      PDF    收藏
Using linker peptide LP4/2A for multiple gene transformation is considered to be an effective method to stack or pyramid several traits in plants. Bacillus thuringiensis (Bt) cry gene and epsps (5-enolpyruvylshikimate-3-phosphate synthase) gene are two important genes for culturing pest-resistant and glyphosate-tolerant crops. We used linker peptide LP4/2A to connect the Bt cry1Ah gene with the 2mG2-epsps gene and combined the wide-used manA gene as a selective marker to construct one coordinated expression vector called p2EPUHLAGN. The expression vector was transferred into maize by Agrobacterium tumefaciens-mediated transformation, and 60 plants were obtained, 40% of which were positive transformants. Molecular detection demonstrated that the two genes in the fusion vector were expressed simultaneously and spliced correctly in translation processing; meanwhile bioassay detection proved the transgenic maize had preferable pest resistance and glyphosate tolerance. Therefore, linker peptide LP4/2A provided a simple and reliable strategy for producing gene stacking in maize and the result showed that the fusion gene transformation system of LP4/2A was feasible in monocot plants.
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16. The Minimal Active Fragment of the Cry1Ai Toxin is Located Between 36I and 605I
ZHOU Zi-shan, LIN Hui-yan, LI Ying, SHU Chang-long, SONG Fu-ping , ZHANG Jie
Journal of Integrative Agriculture    2014, 13 (5): 1036-1042.   DOI: 10.1016/S2095-3119(13)60532-8
摘要1894)      PDF    收藏
The novel cry1Ai gene that cloned from Bacillus thuringiensis strain SC6H8 encoded a protein exhibiting strong toxicity against Plutella xylostella and Chilo suppressalis in our previous study. Using the available information for the active fragments of other Cry toxins, eight truncated fragments were constructed to identify the minimal active fragment of Cry1Ai. All truncated fragments were expressed in Escherichia coli strain BL21 (DE3), and the insecticidal activity against 2nd- instar P. xylostella larvae was assessed using full-length Cry1Ai as a positive control. The results indicate that the minimal active fragment of the Cry1Ai toxin against P. xylostella is located between amino acid residues 36I and 605I, which is smaller than the regions previously reported for Cry1A. The first two amino acids (34T and 35P) on helix α-1 and whole helix α-2 of domain I and sheet β-32 of domain III are necessary for Cry1Ai toxin to keep its toxicity against P. xylostella.
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17. Acquisition of Insect-Resistant Transgenic Maize Harboring a Truncated cry1Ah Gene via Agrobacterium-Mediated Transformation
LI Xiu-ying, LANG Zhi-hong, ZHANG Jie, HE Kang-lai, ZHU Li , HUANG Da-fang
Journal of Integrative Agriculture    2014, 13 (5): 937-944.   DOI: 10.1016/S2095-3119(13)60531-6
摘要2075)      PDF    收藏
A novel insecticidal gene cry1Ah was cloned from Bacillus thuringiensis isolate BT8 previously for plant genetic engineering improvement. Truncated active Cry1Ah toxin has a toxicity level similar to that of the full-length Cry1Ah toxin. In this study, plant expression vector pMhGM harboring truncated cry1Ah gene was transformed into maize (Zea mays L.) immature embryos by Agrobacterium tumefaciens-mediated transformation at which maize alcohol dehydrogenase matrix attachment regions (madMARs) were incorporated on both sides of the gene expression cassette to improve gene expression. A total of 23 PCR positive events were obtained with a transformation efficiency of 5% around. Bioassay results showed that events 1-4 and 1-5 exhibited enhanced resistance to the Asian corn borer (Ostrinia furnacalis). These two events were further confirmed by molecular analysis. Southern blot suggested that a single copy of the cry1Ah gene was successfully integrated into the maize genome. Western blot and ELISA showed that the foreign gene cry1Ah was expressed stably at high level in maize and could be inherited stably over generations. The results of a bioassay of T1-T4 transgenic maize plants indicated that the transgenic plants were highly toxic to the Asian corn borer and their resistance could be inherited stably from generation to generation. Thus, events 1-4 and 1-5 are good candidates for the breeding of insect-resistant maize.
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18. The Influence of Bt-Transgenic Maize Pollen on the Bacterial Diversity in the Midgut of Chinese Honeybees, Apis cerana cerana
JIANG Wei-yu, GENG Li-li, DAI Ping-li, LANG Zhi-hong, SHU Chang-long, LIN Yi, ZHOU Ting, SONG Fu-ping , ZHANG Jie
Journal of Integrative Agriculture    2013, 12 (3): 474-482.   DOI: 10.1016/S2095-3119(13)60248-8
摘要1377)      PDF    收藏
Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the Chinese honeybee, Apis cerana cerana, was assessed. Honeybees were fed with Bt-transgenic maize pollen, non-transgenic near isoline pollen, linear cry1Ah gene (800 ng mL-1) and supercoiled plasmid DNA (800 ng mL-1) under laboratory conditions. The DGGE profile showed that the number of DGGE bands varied from 10.7 to 14.7 per sample, and the Shannon’s index ranged from 0.85 to 1.00. The similarity calculated by PAST was mostly above 92%, indicating no obvious changes among treatments or within replicates. 14 bacterial strains affiliated with Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were isolated and characterized on media under aerobic and anaerobic conditions. These results demonstrated that transgenic cry1Ah maize pollen did not induce significant changes of the honeybee gut bacterial community composition under laboratory conditions.
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19. Application of Mulching Materials of Rainfall Harvesting System for Improving Soil Water and Corn Growth in Northwest of China
HAN Juan, JIA Zhi-kuan, HAN Qing-fang , ZHANG Jie
Journal of Integrative Agriculture    2013, 12 (10): 1712-1721.   DOI: 10.1016/S2095-3119(13)60342-1
摘要1912)      PDF    收藏
The ridge and furrow rainfall harvesting (RFRH) system is used for dryland crop production in northwest of China. To determine the effects of RFRH using different mulching materials on corn growth and water use efficiency (WUE), a field experiment was conducted during 2008-2010 at the Heyang Dryland Experimental Station, China. Four treatments were used in the study. Furrows received uncovered mulching in all RFRH treatments whereas ridges were mulched with plastic film (PF), biodegradable film (BF) or liquid film (LF). A conventional flat field without mulching was used as the control (CK). The results indicated that the average soil water storage at depths of 0-200 cm were 8.2 and 7.3%, respectively higher with PF and BF than with CK. However, LF improved soil water storage during the early growth stage of the crop. Compared with CK, the corn yields with PF and BF were increased by 20.4 and 19.4%, respectively, and WUE with each treatment increased by 23.3 and 21.7%, respectively. There were no significant differences in corn yield or WUE with the PF and BF treatments. The net income was the highest with PF, followed by BF, and the 3-yr average net incomes with these treatments were increased by 2 559 and 2 430 CNY ha-1, respectively, compared with CK. BF and PF had similar effects in enhancing the soil water content, crop yield and net income. Therefore, it can be concluded that biodegradable film may be a sustainable ecological alternative to plastic film for use in the RFRH system in northwest of China.
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20. 胁迫调控因子FgWhi2和磷酸酶FgPsr1在调控禾谷镰孢菌次生代谢物生物合成和对杀菌剂的响应中具有至关重要的作用
Zhang Jie, Gao Han, Ren Fuhao, Zhou Zehua, Wu Huan, Zhao Huahua, Zhang Lu, Zhou Mingguo, Duan Yabing
Journal of Integrative Agriculture    DOI: 10.1016/j.jia.2024.01.003
录用日期: 2023-12-19