Growth promotion effect of Pichia guilliermondii in chilli and biocontrol potential of Hanseniaspora uvarum against Colletotrichum capsici causing fruit rot

In the present investigation, we have attempted to identify the potential two epiphytic yeast strains for growth promotion and management of chilli fruit rot. Seed treatment with Pichia guilliermondii showed increased seedling vigour index (55%), fresh weight (96%) and dry weight (45%) over untreated control. Furthermore, P. guilliermondii showed higher root colonisation ability, indole-3-acetic acid (IAA) production and phosphate solubilisation ability. On the other hand, seedling dip with Hanseniaspora uvarum induced higher levels of defence-related compounds in chilli seedlings challenge-inoculated with Colletotrichum capsici under glasshouse conditions. Among the different media tested, higher biomass of P. guilliermondii and H. uvarum was obtained in pine juice broth and sugarcane juice broth, respectively. Glycerol buffer formulation showed viability (>70%) of P. guilliermondii up to 4 months and H. uvarum up to 9 months when stored at ambient conditions. Seedling dip and foliar sprays with H. uvarum showed 37– 40% reduction in chilli fruit rot incidence under field conditions. It also showed higher (cumulative) accumulation of defence-related compounds in chilli leaves and ripe fruits under field conditions. The results of current investigation indicated a clear difference among the two epiphytic yeast strains. P. guilliermondii was identified as growth promoter of chilli and H. uvarum as antagonist of chilli fruit rot pathogen, C. capsici.     

Role of foliar hemibiotrophic and fruit resistance in anthracnose‐resistant strawberry genotypes for annual hill plasticulture systems

Anthracnose fruit rot (AFR) caused by Colletotrichum acutatum is one of the most devastating diseases of strawberry (Fragaria × ananassa) in plasticulture systems in the Southeast US. Host resistance offers the best option to limit crop losses in both nursery and fruiting fields. To evaluate levels of anthracnose resistance and elucidate the role of foliar and fruit resistance in overall field AFR resistance, we tested 14 strawberry genotypes including numerous selections from the North Carolina State University strawberry breeding programme. Inoculations of plug plants with three representative C. acutatum isolates prior to field‐set indicated that the commercially standard cultivar Chandler was highly susceptible, with an average fruit rot incidence of over 72% the following spring. In contrast, breeding lines such as NC C99‐13 and NC C02‐63 showed superior resistance with AFR incidence values of 23.6% and 11.1%, respectively, and showed superior marketable yields. An average hemibiotrophic infection (HI) severity on foliage (percent leaf area covered with acervular growth after paraquat treatment and incubation) did not correlate (r = 0.57) well with in vitro AFR severity on detached fruit, indicating different mechanisms may be operative for resistance to foliar HI and fruit rot resistance. Multiple regression analysis indicated that in vitro fruit rot resistance expressed by lesion diameter and severity of foliar hemibiotrophic infections may be utilized to predict field AFR incidence. Strawberry genotypes bred for resistance against both fruit rot and foliar HI could be effectively selected by using rank‐sum classification methods and this process offers an effective strategy to advance selections for superior AFR field resistance.     

Hemp sesbania (Sesbania exaltata) control in rice (Oryza sativa) with the bioherbicidal fungus Colletotrichum gloeosporioides f. sp. aeschynomene formulated in an invert emulsion

In greenhouse and field experiments, an invert emulsion (MSG 8.25) was tested with dried, formulated spores of the bioherbicidal fungus Colletotrichum gloeosporioides f. sp. aeschynomene, a highly virulent pathogen of the leguminous weed Aeschynomene virginica (northern jointvetch), but considered ‘immune’ against another more serious leguminous weed, Sesbania exaltata (hemp sesbania). A 1:1 (v/v) fungus/invert emulsion mixture resulted in 100% infection and mortality of inoculated hemp sesbania seedlings over a 21-day period under greenhouse conditions. In replicated field tests of the fungus/invert formulation conducted in Stuttgart, AR, and Stoneville, MS, hemp sesbania was controlled 85 and 90%, respectively. These results suggest that this invert emulsion expands the host range of C. gloeosporioides f. sp. aeschynomene, with a concomitant improvement of the bioherbicidal potential of this pathogen.     

Rapid Detection of Colletotrichum lagenarium,Causal Agent of Anthracnose of Cucurbitaceous Crops,by PCR and Real‐time PCR

Rapid and accurate polymerase chain reaction (PCR) and real‐time PCR methods were developed for the detection of Colletotrichum lagenarium, the causal agent of anthracnose, in tissues of squash (Cucurbita moschata), watermelon (Citrullus lanatus), cucumber (Cucumis sativus) and muskmelon (Cucumis melo). PCR assays amplified different internal transcribed spacer sequences from C. lagenarium, so effectively detected this pathogen in infected tissues. PCR analysis with the primer co‐m‐337F1/R1 was able to differentiate C. lagenarium from other fungal pathogens, including Colletotrichum spp., Fusarium spp., Alternaria spp. and Didymella spp. An optimized real‐time PCR assay was developed to detect and monitor C. lagenarium in both infected plant tissues and soil samples. The sensitivity of real‐time PCR can detect down to 1 pg of DNA. Thus, PCR‐based analysis is a useful technique for rapid detection and diagnosis of C. lagenarium in infected plants or infested soils.     

京津水源区小流域土壤侵蚀空间模拟

针对京津水源区生态环境脆弱、水土流失空间分异大、突发性强等问题,以河北省平泉县东北沟典型小流域为例,采用GIS技术与CSLE模型对其土壤侵蚀进行二维空间模拟及侵蚀规律研究,并对模型模拟精度进行验证.结果表明:模型确定性系数>0.85,模拟结果可信度高;小流域侵蚀模数为2359.24t·km-2·a-1,属于轻度侵蚀,侵蚀主要源于15°以上坡面,占侵蚀总量的92.1%;土壤侵蚀模数随坡度先逐渐增加,当坡度>45°时,土壤侵蚀模数有减小的趋势;土壤侵蚀模数空间异质性显著,坡耕地侵蚀模数最大,为6112.90tkm-2·a-1;荒草地占总面积33.2%,侵蚀量却达总量的74.9% ;59.3%的侵蚀来自阳坡和半阳坡,而阴坡半阴坡仅占流域侵蚀量的16.2%.研究为利用CSLE模型对小流域土壤侵蚀进行空间模拟提供技术范例,为京津水源区土壤侵蚀规律研究及水土保持生态建设提供科学参考.     

云南芒果采后炭疽病病原菌的鉴定及室内生防菌的筛选

随着对云南芒果需求量的增加,其种植面积日趋扩大,并不断引入新品种,这也导致云南省芒果炭疽病害发生日趋加重。为了有针对性地开展芒果炭疽病的生物防治,采用组织块分离法分离芒果采后炭疽病病原菌,通过形态学观察初步鉴定,并遵循柯赫氏法则对病原菌进行验证。随后,利用rDNA-ITS序列和系统发育树分析明确病原菌的分类学地位。最后,采用5种生防细菌对病原菌进行拮抗试验。通过对芒果采后炭疽病病原菌进行分离鉴定,确定胶孢炭疽菌(Colletotrichum gloeosporioides)是引起云南芒果采后炭疽病的病原菌,该菌株内转录间隔区(internal transcribed spacer,ITS)序列长度为536 bp,登录号为MH744668;5株生防细菌对芒果炭疽病病原菌都有一定的抑菌作用,具有较好的生防开发潜能,其中抗生素溶杆菌L-44的抑菌效果最好,抑制率达53.7%。研究结果为云南省芒果采后病害的生物防治提供了新的思路。     

基于CRISPR-Cas9技术构建橡胶树胶孢炭疽菌的基因敲除系统

[背景]CRISPR-Cas9基因组编辑技术为病原真菌的基因敲除、敲入及定点编辑提供了新的思路。[目的]建立适用于橡胶树胶孢炭疽菌的CRISPR-Cas9基因敲除系统。[方法]通过大肠杆菌原核表达系统合成含有细胞核定位信号的Cas9蛋白;以URA5为靶标基因,预测该基因中Cas9的切割位点,并在体外转录合成相应的SgRNA;体外构建Cas9-SgRNA复合体,并将该复合体转入橡胶树胶孢炭疽菌原生质体;通过表型筛选及测序鉴定,筛选URA5的敲除突变体菌株。[结果]体外表达的Cas9蛋白与SgRNA能够形成复合体,并在体外对目标基因URA5的DNA序列进行切割;Cas9-SgRNA复合体能够成功转入橡胶树胶孢炭疽菌原生质体,并完成对URA5的敲除;敲除突变株表现出尿嘧啶缺陷表现型。[结论]建立了适用于橡胶树胶孢炭疽菌的基因敲除系统。     

Morphological and molecular identification of Colletotrichum siamense,a novel leaf pathogen associated with Sterculia lanceolata recorded in China

Sterculia lanceolata, an important tropical woody plant, has high ornamental and medicinal value. To our knowledge, only brown root disease in this plant has been reported. In Nanning, Guangxi, China, an outbreak of leaf blight disease was observed on S. lanceolata in June 2019, with the leaf infection rate ranging from 80% to 100%. The disease seriously affected the leaves of trees and caused economic loss. Eight isolates were recovered from the infected leaves of different trees, and the pathogenicity was then determined by the methods of mycelial disc and conidial suspension, fulfilling Koch's postulates. According to the morphological and molecular biological characteristics of isolates, the pathogen causing leaf blight on S. lanceolata was identified as Colletotrichum siamense. Accurate identification of the pathogen provides a reliable basis for the control of the disease.     

Best linear unbiased prediction of host-range of the facultative parasite Colletotrichum gloeosporioides f. sp. salsolae, a potential biological control agent of Russian thistle

Russian thistle or tumbleweed (Salsola tragus L.) is an introduced invasive weed in N. America. It is widely distributed in the US and is a target of biological control efforts.The fungus Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. in Penz. f. sp. salsolae (CGS) is a facultative parasite under evaluation for classical biological control of this weed. Host-range tests were conducted with CGS in quarantine to determine whether the fungus is safe to release in N. America. Ninetytwo accessions were analyzed from 19 families: Aizoaceae, Alliaceae, Amaranthaceae, Apiaceae, Asteraceae, Brassicaceae, Cactaceae, Campanulaceae, Chenopodiaceae, Cucurbitaceae, Cupressaceae, Fabaceae, Malvaceae, Nyctaginaceae, Phytolaccaceae, Poaceae, Polygonaceae, Sarcobataceae, and Solanaceae and 10 tribes within the Chenopodiaceae: Atripliceae, Beteae, Camphorosmeae, Chenopodieae, Corispermeae, Halopepideae, Polycnemeae, Salicornieae, Salsoleae, and Suaedeae. These included 62 genera and 120 species. To facilitate interpretation of results, disease reaction data were combined with a relationship matrix derived from internal transcribed spacer DNA sequences and analyzed with mixed model equations to produce Best Linear Unbiased Predictors (BLUPs) for each species. Twenty-nine species (30 accessions) from seven closely-related Chenopodiaceae tribes had significant levels of disease severity as indicated by BLUPs, compared to six species determined to be susceptible with least squares means estimates. The 29 susceptible species were: 1 from Atripliceae, 4 from Camphorosmeae, 1 from Halopepideae, 2 from Polycnemeae, 6 from Salicornieae, 8 from Salsolae, and 7 from Suaedeae. Most species in the genus Salsola, which are all introduced and weedy, were very susceptible and damaged by CGS. Statistical comparisons and contrasts of BLUPs indicated that these Salsola species were significantly more susceptible than non-target species, including 15 species from relatives in the closely-related genera Bassia (=Kochia), Nitrophila, Salicornia, Sarcocornia, and Suaeda. Of the 29 susceptible species, 10 native or commercially important species in N. America were identified as needing additional tests to determine the extent of any damage caused by infection.