Biological control serves as a crucial strategy for crop disease management. The biocontrol potential and plant growth-promoting effects of the strain YTBTa14 were investigated. Genetic sequencing confirmed YTBTa14 as Pseudomonas chlororaphis, which exhibited broad-spectrum antifungal activity against multiple pathogens affecting grapevine, apple, cherry, and wheat. YTBTa14 significantly enhanced the growth of wheat and grapevine, specifically increasing wheat seed germination rates and improving root and coleoptile development. In grapevine plant, significant increases in root length, stem length, and fresh weight were observed. The strain demonstrated robust adaptability and stable antagonism under varying sodium chloride (NaCl) concentrations, pH levels, and temperatures. YTBTa14 modulated plant hormone levels, elevating the content of indole-3-acetic acid (IAA), gibberellins (GA), and cytokinins (CTK). Furthermore, it effectively stimulated the production of key plant defense enzymes, including superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT). Pretreatment of grape leaves with YTBTa14 triggered plant cell defense response and upregulated the expression of defense-related genes PR1 (pathogenesis-related protein 1) and PAL1 (phenylalanine ammonia-lyase 1), thereby mitigating the severity of downy mildew disease and inducing systemic resistance. These findings demonstrate that YTBTa14 is a highly promising candidate for development as a multifunctional agricultural biocontrol agent.

CRISPR/Cas12a, a promising gene editing technology, faces limitations due to its requirement for a thymine (T)-rich protospacer adjacent motif (PAM). Despite the development of Cas12a variants with expanded PAM profiles, many genomic loci, especially those with guanine-cytosine (GC)-rich PAMs, have remained inaccessible. This study develops a small RNA toxin-aided strategy to evolve ErCas12a for targeting GC-rich PAMs, resulting in the creation of enhanced ErCas12a (enErCas12a). EnErCas12a demonstrates the ability to recognize GC-rich PAMs and target five times more PAM sequences than the wild-type ErCas12a. Furthermore, enErCas12a achieves efficient gene editing in both bacterial and mammalian cells at various sites with non-canonical PAMs, including GC-rich PAMs such as GCCC, CGCC, and GGCC, which are inaccessible to previous Cas12a variants. Moreover, enErCas12a effectively targets PAM sequences with a GC content exceeding 75% in mammalian cells, providing a valuable alternative to the existing Cas12a toolkit. Importantly, enErCas12a maintains high specificity at targets with canonical PAMs, while also demonstrating enhanced specificity at targets with non-canonical PAMs. Collectively, this work establishes enErCas12a as a promising tool for gene editing in both eukaryotes and prokaryotes.

Leaf spot, a major apple disease, manifests in diverse symptoms. In this study, the pathogen was isolated from diseased ‘Yanfu 3’ apple leaves in Yantai, Shandong Province, and identified as Neopestalotiopsis clavispora through morphological observation, molecular identification, and multi-gene (ITS, TEF1α, and TUB2) phylogenetic analysis. Three isolates (YTNK01, YTNK02, and YTNK03) were selected for pathogenicity tests to verify Koch’s postulates. To our knowledge, this is the first report of N. clavispora being responsible for apple leaf spots in China, and the disease has been named ‘apple Neopestalotiopsis leaf spot’. Additionally, N. clavispora was found to infect crabapple, sweet cherry, grape, peach, and pear under laboratory conditions, indicating that these fruit trees may be potential hosts for N. clavispora in the field. The in vitro toxicity of ten fungicides to the pathogen was assessed using the mycelial growth rate method. All ten fungicides were effective in inhibiting the growth of N. clavispora. Among them, those based on pylocyanonitrile, propiconazole, pyraclostrobin, tebuconazole, diphenoxazole, and osthole showed higher toxicity to N. clavispora, with EC50 values of 0.11, 0.41, 0.47, 1.32, 1.85, and 3.82 µg/mL, respectively. These fungicides could be used as alternatives to prevent this disease in production. Overall, these findings provide valuable insights into the characteristics of N. clavispora causing apple leaf spot and are crucial for developing effective management strategies.

苹果病虫害绿色防控关系到果品品质和质量安全,对促进苹果产业绿色高质量发展具有重要意义.综述了苹果病虫害绿色防控技术及研究进展,主要从监测预警、农药减量、物理防治、生物防治、生态调控等方面进行归纳总结,提出了苹果病虫害绿色防控存在的问题及今后的工作方向.

为明确宛氏拟青霉乙醇提取物ZNC(以下简称ZNC)在花生上的应用效果,于烟台市进行了田间试验.结果表明,施用ZNC能明显促进花生生长,用ZNC 40 mL拌花生种100 kg播种后,苗期和花期再进行2次叶面喷雾处理表现出效果最为显著,株高、叶面积和叶绿素含量较清水对照分别增加17.06 cm、5.88 cm2和1.61 mg/g.在诱导抗病性上,ZNC对花生叶斑病的最高防效达38.62％,显著提高了抗病性.在产量方面,施ZNC处理的花生产量增加6.80％～26.30％.总之,施用ZNC能够促进花生生长、提高产量、增强抗病性,可以在本地花生生产中推广应用.

Apple Alternaria blotch disease, caused by Alternaria alternata (Fr.) Keissl, is one of the most famous leaf diseases. When the disease is prevalent, it causes leaf abscission and influences the formation of flower buds and photosynthesis. Therefore, a simple, rapid, high-specificity and sensitivity method for monitoring infected leaves at early developmental stages is urgently needed, so that the occurrence and expansion of A. alternata can be controlled in time. In our research, a rapid, specific and efficient loop-mediated isothermal amplification (LAMP) method was developed to detect A. alternata within 60 min. Six primers of LAMP detection can only specifically amplify the aapg-1 gene in A. alternata but not in four other important fungi in apples. The aapg-1 gene encodes endopolygalacturonase in A. alternata, and there are significant differences among different species. Thus, it was applied as the target for LAMP primers. Compared to conventional PCR detection, our LAMP method had the same sensitivity as that of detecting as little as 1 fg of pure genomic DNA of A. alternata. When leaves were inoculated with A. alternata conidia, LAMP detected 1 × 102 conidia/mL as the minimum concentration. However, the traditional tissue isolation and identification method only isolated A. alternata from leaves inoculated with 1 × 105 and 1 × 106 conidia/mL, indicating that the LAMP method was more sensitive than the traditional tissue isolation and identification method for A. alternata before symptoms. Further tests also indicated that LAMP detection was more accurate and sensitive than the traditional tissue isolation and identification method for A. alternata in leaves with the Alternaria blotch symptom collected from the field. Our results showed that the LAMP-targeting the aapg-1 gene has the advantages of high sensitivity, specificity and simplicity and can be used for rapid detection and early monitoring of A. alternata in the field. LAMP is instructive for us to effectively prevent and control apple Alternaria blotch disease.

橘小实蝇是为害果蔬的重大害虫,以成虫诱杀为主要作用方式的食诱技术是防治该害虫的重要方法.综述了目前橘小实蝇食诱剂的主要类型及研究进展,并对其在生产中搭配诱捕器、光波、化学物质等主要应用技术进行了归纳,提出了橘小实蝇食诱技术应用中存在的问题和今后的研究方向.

明确病虫害发生情况是果园农药减量防控的基础.近年来,随着矮化密植技术的推广,苹果园生态环境、树形、病虫害均发生了很大变化,传统的病虫害调查方法已不再适用.对新型矮化密植苹果园,根据叶片病虫害的发生特点,在传统调查取样方法的基础上进行了修改与完善,减少单株取样量,增加株调查数量,更加科学地反映苹果园叶片病虫害的发生情况.

为明确新型杀虫剂5％双丙环虫酯可分散液剂和17％氟吡呋喃酮可溶液剂对桃蚜的田间防治效果,2019—2020年于山东省烟台市桃园开展了田间药效试验.结果表明,5％双丙环虫酯可分散液剂3000～5000倍液、17％氟吡呋喃酮可溶液剂4000～6000倍液对桃蚜的防效优于对照药剂70％吡虫啉水分散粒剂12000倍液.5％双丙环虫酯可分散液剂、17％氟吡呋喃酮可溶液剂可作为防治桃蚜的高效药剂在生产中与常规药剂交替使用.

为明确42.8％氟吡菌酰胺·肟菌酯悬浮剂对葡萄白腐病、炭疽病、灰霉病的综合防治效果,2017—2018年于山东烟台进行了田间药效试验.结果表明,42.8％氟吡菌酰胺·肟菌酯悬浮剂125.0～187.5 mg/kg对葡萄白腐病防效在91.97％以上,对炭疽病在87.34％以上,其125.0～250.0 mg/kg对葡萄灰霉病防效在86％以上.42.8％氟吡菌酰胺·肟菌酯悬浮剂是兼防葡萄白腐病、炭疽病、灰霉病3种果实病害的高效药剂.

With the rapid development of electrolytic manganese industry, the environmental problems caused by the storage of electrolytic manganese slag are particularly prominent. It not only occupies land resources, but also easily causes heavy metal pollution in soil, surface water and groundwater. Therefore, it is necessary to treat electrolytic manganese slag safely and effectively. The paper mainly studies the solidification/stabilization of electrolytic manganese slag and its environmental safety for road filling, in order to open up a new way of harmless and resource utilization of electrolytic manganese slag. In this paper, lime and fly ash were used as stabilizers and cement was used as curing agent to stabilize manganese slag, and the stabilization effect of Mn and Pb in manganese slag was studied. The stabilization effect of manganese (Mn) and lead (Pb) in manganese slag was studied. The results show that the dosage of stabilizer quicklime is 2.5%, fly ash is 3%, and the dosage of solidifying agent cement is 12%, the solidification/stabilization effect is the best compared with other ratios, then the leaching concentrations of Mn and Pb meets the requirements of China’s surface water environmental quality standards for category III water sources, which can be used as domestic water after treatment. Under the optimal ratio of stabilization effect, the compressive strength and slump are 13.8 MPa and 50 mm, respectively. The research results of the paper can provide a new way for the harmless treatment of manganese slag and the resource utilization of new materials.

为明确不同葡萄品种霜霉病的田间发生流行规律,连续4年田间定点调查无核红宝石、玫瑰香、蛇龙珠葡萄霜霉病的初始发生时间及规律.调查表明,葡萄霜霉病在无核红宝石、玫瑰香、蛇龙珠上的初发时期及危害程度不同.无核红宝石于7月中、下旬始见葡萄霜霉病斑,比玫瑰香早7 d左右,比蛇龙珠早10 d左右;霜霉病病情指数,无核红宝石同期均显著高于玫瑰香和蛇龙珠;无核红宝石发病高峰期时间最早,在9月20日前达到高峰,比玫瑰香早6 d左右,比蛇龙珠早10 d左右.

为明确山东省胶东地区葡萄白腐病菌Coniella diplodiella对抑霉唑的敏感性及抑霉唑与常规杀菌剂之间的交互抗性,采用菌丝生长速率法测定葡萄白腐病菌对抑霉唑等杀菌剂的敏感性,并通过分析抑霉唑与戊唑醇、吡唑醚菌酯、福美双、多菌灵、代森锰锌毒力的相关性,判断抑霉唑与各药剂之间是否存在交互抗性.结果 表明,供试69株葡萄白腐病菌菌株对抑霉唑的EC50在0.13～55.53 μg/mL之间,最高值与最低值相差427.15倍;其频数分布图呈多峰曲线,第一主峰内EC50平均值为6.34 μg/mL,可作为胶东地区葡萄白腐病菌对抑霉唑的敏感基线.与敏感基线相比,田间已出现抑霉唑低抗菌株,占检测总株数的8.70％,未检测到中、高抗菌株.选择3个抗性菌株连续继代培养10代后,其对抑霉唑的敏感性明显提高,说明其抗药性不能稳定遗传.抑霉唑EC50对数值与戊唑醇、吡唑醚菌酯、福美双、多菌灵、代森锰锌的EC50对数值之间的相关系数分别为0.799、-0.143、-0.089、-0.268和0.159,说明抑霉唑与戊唑醇存在一定的交互抗性,但与其他4种药剂之间不存在交互抗性.表明抑霉唑可用于胶东地区田间葡萄白腐病的有效防控.

为筛选防治葡萄炭疽病安全、高效的生物药剂,采用菌丝生长速率法测定5种生物药剂对葡萄炭疽病菌的毒力,选取其中抑菌活性较好的4种药剂开展田间防效试验.室内毒力测定结果表明,丁子香酚、苦参碱、中生菌素、乙蒜素的EC50分别为0.50、20.03、22.05、33.04mg/L,均低于对照药剂代森锰锌的EC50值42.66mg/L,抑菌效果较好,而多抗霉素的EC50为3 420.15 mg/L,抑菌效果差.田间药效试验结果表明,0.3％丁子香酚可溶性液剂1 000-1 250倍液对葡萄炭疽病的防效最佳,为91.41％～93.49％,显著优于对照药剂80％代森锰锌可湿性粉剂800倍液,0.3％丁子香酚可溶性液剂1 500倍液、80％乙蒜素乳油1 000-1 500倍液防效虽低于对照药剂,但也均在80％以上,可作为与常规化学药剂轮换使用防控炭疽病的有效生物药剂进行推广应用.

[目的]明确鸡粪沼液与化学农药噻虫胺减量混用对韭菜迟眼蕈蚊的防治效果.[方法]分别采用胃毒触杀法和灌根法进行室内毒力测定和田间药效试验.[结果]鸡粪沼液稀释50倍与1 mg/L噻虫胺减量10％～40％混用,对韭菜迟眼蕈蚊3龄幼虫的毒力与1 mg/L噻虫胺差异不显著.鸡粪沼液50～100倍在药剂减少20％的情况下,桶混制剂对韭菜迟眼蕈蚊的田间防效与噻虫胺常规剂量相当.[结论]在使用噻虫胺防治韭菜迟眼蕈蚊时,建议鸡粪沼液稀释50～100倍,与噻虫胺减量20％混用.

[目的]明确新型杀菌剂10％四唑吡氨酯悬浮剂对黄瓜霜霉病的防治效果,为生产上应用提供科学依据.[方法]采用喷雾法于2016-2017年在山东省烟台市进行了田间药效试验.[结果]10％四唑吡氨酯悬浮剂对黄瓜霜霉病防治效果良好,第3次施药后14 d,30～90 g a.i./hm2处理防效仍在88.62％以上,优于对照药剂50％烯酰吗啉水分散粒剂(225 g a.i./hm2).[结论]10％四唑吡氨酯悬浮剂是一种防治黄瓜霜霉病较为理想的新型药剂,值得在黄瓜生产中推广应用.

为了明确葡萄园绿盲蝽田间种群的抗药性,采用试管药膜法测定了噻虫胺、高效氯氰菊酯、毒死蜱、阿维菌素、吡虫啉对山东省烟台市不同地区绿盲蝽种群的室内毒力.结果 表明:噻虫胺对烟台地区绿盲蝽的毒力最高,阿维菌素次之,再次为吡虫啉和高效氯氰菊酯,毒死蜱对烟台地区绿盲蝽的毒力最低;烟台市不同地区绿盲蝽种群对供试药剂的敏感性差异较小,而福山地区的绿盲蝽种群对各供试药剂的敏感性最高,莱山地区的绿盲蝽种群对各供试药剂的敏感性最低.

为建立快捷、灵敏检测苹果轮纹病菌Botryosphaeria dothidea的环介导等温扩增(loop-mediated isothermal amplification,LAMP)检测方法,以其内转录间隔区ITS序列为靶标,设计6条LAMP引物,对其特异性进行检测,优化反应条件并建立苹果轮纹病菌的LAMP检测方法.引物特异性检测结果表明,2株苹果轮纹病菌反应结果呈绿色为阳性,而其它3株对照菌株反应结果呈橙色为阴性,表明了LAMP检测引物的高特异性.优化后的LAMP最佳反应条件:温度65℃、扩增时间60 min、FIP/BIP、F3/B3、LF/LB引物终浓度分别为1.0、0.25、0.5 μmol/L.LAMP检测方法对苹果轮纹病菌DNA的检测灵敏度达到了100 ag/μL,是常规PCR检测灵敏度的100倍.田间疑似轮纹病组织检测结果发现LAMP方法对苹果轮纹病菌的检出率高达68％,而传统分离鉴定方法的检出率仅为24％.表明所建立的苹果轮纹病菌LAMP快速检测方法简便快捷、特异性好、灵敏度高,尤其适用于基层植保机构对于苹果轮纹病菌的田间快速检测.

The endophytic Bacillus amyloliquefaciens YTB1407 was previously reported to promote the growth of sweet potato (Ipomoea batatas cv. Yanshu 25). Here, we demonstrate in both in vitro and pot trial assays that pre-treatment with YTB1407 suspension could enhance resistance against root rot disease and black rot disease, caused by Fusarium solani Mart. Sacc. f. sp. batatas McClure and Ceratocystis fimbriata Ell. & Halst on sweet potato, respectively. When seedlings were infected with fungal pathogens at 10 days post irrigation, pre-treatment with YTB1407 suspension decreased these pathogens and YTB1407 bacterial biomass in sweet potato roots. The pre-treatment activated the expression of salicylic acid (SA)-responsive PR-1 gene, raised SA content, and reduced hydrogen peroxide (H2O2) in the host to resist F. solani, while it enhanced the expression levels of SA-responsive NPR1 and PR1 genes and increased SA content to resist C. fimbriata. The disease resistance control effect initiated by pre-treatment with YTB1407 for root rot pathogen (F. solani) was better than for black rot pathogen (C. fimbriata). The results indicated that Bacillus amyloliquefaciens YTB1407 played a pivotal role in enhancing resistance to two fungi pathogens in sweet potato, through production of some antifungal metabolites to decrease infection in the early stage as well as induction of SA-dependent systemic resistance.

针对山东产区套袋苹果的常见病虫害,设计了病虫害综合防控“164”农药减施模式,即1个重点生态防控期、6个化学防治关键期和4个病虫重点监控期.按“164”减药模式制定适用不同地区和采用不同农药公司药剂种类的具体防控方案,在栖霞、蓬莱等地的果园连续试验2年.结果 表明:试验园全年用药9～ 14药次,与当地常规管理的对照园相比,化学农药使用量减少50％以上;试验园的病虫叶率和病虫果率都控制在5％以内,果园内无落叶现象;病虫防治效果、果实的产量和品质、优质商品果率等项指标,与当地常规管理的对照园无明显差异.