tree nuts

树坚果
  • 文章类型: Journal Article
    中国核桃的生产和收获面积居世界第一。目前,病原体感染引起的核桃健康状况不佳和产量低令人担忧。2022年,聊城市4个核桃苗圃(0.08~0.23公顷)幼苗上出现严重的核桃叶斑病,山东,中国,平均发生率为48.6%(香陵品种的34.6%至65.3%)。从8月到10月,叶斑主要出现在小叶的边缘,偶尔在静脉之间。病变最初柔软腐烂,然后是浅棕色,圆形到半圆形。随后,相邻的病变融合,小叶和整张小叶的边缘显示褐变和枯萎的症状。对于病原体分离,收集其中一个苗圃的五个具有代表性症状的传单,并用浸入75%酒精的无菌脱脂棉擦拭三次,并用蒸馏水洗涤。切除病变与健康组织交界处的小叶碎片,用无菌砂浆压碎,并在少量蒸馏水中浸泡10分钟。将患病组织悬浮液在具有无菌接种环的营养琼脂培养基(NA)上划线,并在28°C下孵育24至72小时。将获得的细菌菌落在NA上进一步培养。纯化的菌落形状均匀,圆形,黄色,一个被举起的,有光泽的表面和光滑的边缘。分离株为革兰氏阴性,电子显微镜分析表明,病原体为短棒(0.35至0.52×0.90至1.24μm,平均值=0.44±0.05×1.08±0.11μm,n=25)。对于细菌种类鉴定,单菌落培养进行基因组DNA提取和16SrRNA的基因扩增和测序,rpoD,还有gyrb.通用引物27F/1492R(Lane1991)用于扩增16SrRNA基因,特异性引物70F/70R和UP-1E/APrU(Yamamoto等人。2000)用于扩增rpoD和gyrB基因,分别。在BLAST分析中,该分离株的16SrRNA序列(GenBankOR195734)与假单胞菌菌株IAM1568T(AM262973.1)具有99%的相似性(1409/1410bp),rpoD(OR709708)和gyrB(OR709707)序列显示与鱼群品系LMG7040T的rpoD(707/717bp;FN554494.1)和gyrB(787/801bp;FN554210.1)>98%的同一性。基于以上结果,分离的细菌被鉴定为P.oryzihibitans。对于致病性测试,来自10个两岁的盆栽核桃幼苗的健康小叶(cv。香玲)作为接种材料。用0.4毫米的无菌接种针刺穿小叶,每个小叶上间隔约5mm的三个小孔用一块无菌棉覆盖。将107CFU/ml的细菌悬浮液(1ml)铺在棉花上,并用塑料薄膜包裹24小时。水用作阴性对照。接种进行五次。植物在室外生长,日平均温度为22°C,相对湿度超过45%。接种后两天,这种疾病开始在小叶中发展,症状与在野外观察到的症状相似。相比之下,对照植物保持健康无症状。从接种的核桃植物中重新分离出细菌,分离株的形态和16SrRNA基因序列与原始菌株相同。自从在1970年代被发现为机会性人类致病菌以来(Keikha等人。2019),P.oryzihabitans也被证明会引起某些植物病害,例如水稻上的穗枯病和谷物变色(Hou等人。2020),花椒上的果实黑腐病(Liu等人。2021),甜瓜的茎和叶腐病(Li等人。2021)。据我们所知,这是我国首例引起核桃叶斑病的水稻斑点病的报道。由P.oryzihabitans引起的叶斑病可能是核桃种植的威胁,此报告是确定潜在传播和有效控制措施的第一步。
    China ranks first in the production and harvest area of walnut (Juglans regia L.) worldwide. Currently, the poor health and low yield of walnut caused by pathogen infection is of concern. In 2022, severe walnut leaf spot disease was observed on the seedlings of four walnut nurseries (0.08 to 0.23 ha) in Liaocheng, Shandong, China, with an average incidence of 48.6% (from 34.6% to 65.3% on the cultivar Xiangling). From August to October, leaf spots mainly appeared on the edges of the leaflets, and occasionally between veins. The lesions were initially soft and rotten, and then light brown, round to semi-circular. Subsequently, the adjacent lesions fused, and the edges of the leaflets and entire leaflets showed symptoms of browning and wilting. For pathogen isolation, five leaflets with representative symptoms from one of the nurseries were collected and wiped three times with sterile absorbent cotton dipped in 75% alcohol and washed with distilled water. Leaflet pieces at the junction of the lesion and healthy tissues were removed, crushed in a sterile mortar, and soaked in a small amount of distilled water for 10 min. The diseased tissue suspension was streaked on a nutrient agar medium (NA) with a sterile inoculation ring and incubated at 28°C for 24 to 72 h. The bacterial colonies obtained were further cultured on NA. The purified colonies were uniform in shape, round, and yellow, with a raised, shiny surface and smooth margin. The isolates were Gram-negative, and the electron microscope analysis showed that the pathogens were short rods (0.35 to 0.52 × 0.90 to 1.24 μm, average = 0.44 ± 0.05 × 1.08 ± 0.11 μm, n = 25). For bacterial species identification, a single-colony culture was subjected to genomic DNA extraction and gene amplification and sequencing of 16S rRNA, rpoD, and gyrB. The universal primers 27F/1492R (Lane 1991) were used to amplify the 16S rRNA gene and the specific primers 70F/70R and UP-1E/APrU (Yamamoto et al. 2000) were used to amplify the rpoD and gyrB genes, respectively. In the BLAST analysis, the 16S rRNA sequence (GenBank OR195734) of the isolate shared 99% similarity (1409/1410 bp) with Pseudomonas oryzihabitans strain IAM 1568T (AM262973.1), and the rpoD (OR709708) and gyrB (OR709707) sequences showed >98% identity to rpoD (707/717 bp; FN554494.1) and gyrB (787/801 bp; FN554210.1) of P. oryzihabitans strain LMG 7040T. Based on the above results, the isolated bacterium was identified as P. oryzihabitans. For the pathogenicity test, healthy leaflets from 10 two-year-old potted walnut seedlings (cv. Xiangling) were used as inoculation materials. The leaflets were punctured with a sterile inoculation needle of 0.4 mm, and three small holes on each leaflet at an interval of about 5 mm were covered with a piece of sterile cotton. A bacterial suspension (1 ml) at 107 CFU/ml was spread onto the cotton, and wrapped with plastic film for 24 h. Water was used as a negative control. The inoculations were performed five times. Plants were grown outdoors at a daily average temperature of 22°C with relative humidity over 45%. Two days after inoculation, the disease began to develop in the leaflets with similar symptoms to those observed in the field. In contrast, control plants remained healthy and symptomless. Bacteria were reisolated from the inoculated walnut plants, and the morphology and 16S rRNA gene sequences of the isolates were the same as those of the original strains. Since it was discovered as an opportunistic human pathogenic bacterium in the 1970s (Keikha et al. 2019), P. oryzihabitans has also been shown to cause certain plant diseases, such as panicle blight and grain discoloration on rice (Hou et al. 2020), fruit black rot on prickly ash (Liu et al. 2021), and stem and leaf rot on muskmelon (Li et al. 2021). As far as we know, this is the first report of P. oryzihabitans causing walnut leaf spot disease in China. Leaf spot caused by P. oryzihabitans may be a threat to walnut cultivation, and this report of its occurrence is the first step in determining potential spread and effective control measures.
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  • 文章类型: Journal Article
    英国核桃(JuglansregiaL.)是多年生落叶果树,和世界范围内种植的重要经济硬木树种。作为重要的经济作物之一,英国核桃在新疆也广泛种植。2019年9月,在几个果园的英国核桃上观察到树枝溃疡症状,南疆地区的疾病发病率约为15%至40%(79º95\'E,40º37\'N)。分支病变长椭圆形,凹面,从黑色到棕色。受影响的树枝的叶子变成黄色,树枝最终死亡。从果园中被感染的树上收集被感染的树枝。用75%乙醇对溃疡边缘的症状组织进行表面消毒60s,用无菌水冲洗3次,然后在25℃的马铃薯葡萄糖琼脂培养基(PDA)上,在光培养箱中12小时的光周期下孵育7天。从有症状的组织中获得了7种表现出相似形态的真菌分离株。所有的真菌培养物都是粉白色的,松散的,棉质菌丝体,殖民地的下面是浅棕色的。大分生孢子略微弯曲,用一到六个隔片,两端都有点锋利,他们测量22.8至38.5×3.5至6.7μm(27.4±0.6×4.2±0.3μm,n=50)。微分生孢子是椭圆形的,透明玻璃,零到一个隔膜,测量4.5至9.6×1.8至2.3μm(6.8±0.3×2.1±0.1μm,n=50)。根据形态特征,这七个分离株被鉴定为枯萎病镰刀菌物种复合体的成员(Summerell等人。2003).从代表性分离株HSANTUAN2019-1和内部转录间隔区(ITS)区域中提取基因组DNA,用引物对ITS1/ITS4扩增翻译延伸因子1-α(TEF)(White等人。1990)和EF1-F/EF2-R(O'Donnell等人。2010),分别。提交给GenBank的序列(登录号用于ITS的OP271472,用于TEF的OP293104)显示与F.solani(ITS,OL691083[100%];TEF,HE647960[99.86%])。在田间1岁的英国核桃分支上评估了七个分离株的致病性。健康的树枝(40)被消毒的打孔器打伤,然后用等直径的菌丝体PDA塞(每个真菌分离株5个分支)接种。用无菌PDA栓接种五个分支作为阴性对照。接种进行三次。所有处理用新鲜膜包裹3天。接种后22天在所有接种的分支上观察到深棕色坏死病变。对照没有症状。从所有接种的分支中重新分离出病原体,履行科赫的假设。据我们所知,这是F.solani在新疆引起英国核桃树枝溃疡的第一份报告,中国。小枝溃疡病经常引起大量的枝条干裂和死亡。如果忽视疾病控制和预防,种植面积严重影响了英国核桃的生产力。我们的发现将为英国核桃树枝溃疡的预防和管理提供有价值的信息。
    English walnut (Juglans regia L.) is a perennial deciduous fruit tree, and an economically important hardwood tree species cultivated worldwide. As one of the important economic crops, English walnut is also widely cultivated in Xinjiang. In September 2019, twig canker symptoms were observed on English walnut in several orchards, with approximately 15% to 40% disease incidence in southern Xinjiang region (79º95\'E, 40º37\'N). The branch lesions were long oval, concave, and black to brown. Leaves of the affected branches turned yellow and the branches eventually died. Infected twigs were collected from an infected tree in an orchard. Symptomatic tissue from the margins of cankers was surface disinfested with 75% ethanol for 60 s, rinsed 3 times with sterile water, and then incubated on potato dextrose agar medium (PDA) at 25 ℃ under a 12-hr photoperiod in Light incubator for 7 days. Seven fungal isolates showing similar morphology were obtained from the symptomatic tissue. All the fungal cultures had a pink-white color with loose, cottony mycelium, and the underside of the colonies were light brown. Macroconidia were slightly curved, with one to six septa, both ends were slightly sharp, and they measured 22.8 to 38.5 × 3.5 to 6.7 μm (27.4 ± 0.6 × 4.2 ± 0.3 μm, n=50). Microconidia were oval, hyaline, zero to one septa, measuring 4.5 to 9.6 × 1.8 to 2.3 μm (6.8 ± 0.3 × 2.1 ± 0.1 μm, n=50). According to the morphological characteristics, the seven isolates were identified as a member of the Fusarium solani species complex (Summerell et al. 2003). Genomic DNA was extracted from the representative isolate HSANTUAN2019-1, and the internal transcribed spacer (ITS) region, the translation elongation factor 1-alpha (TEF) were amplified with the primer pairs ITS1/ITS4 (White et al. 1990) and EF1-F/EF2-R (O\' Donnell et al. 2010), respectively. The sequences submitted to GenBank (accession nos. OP271472 for ITS, OP293104 for TEF) showed high similarity with the reference sequences of F. solani (ITS, OL691083 [100%]; TEF, HE647960 [99.86%]). Pathogenicity of the seven isolates were assessed on 1-year-old branches of English walnut in the field. Healthy branches (40) were wounded with a sterilized hole punch, and then inoculated with isodiametric mycelial PDA plugs (5 branches per fungal isolate). Five branches were inoculated with sterile PDA plugs as a negative control. The inoculations were performed three times. All treatments were wrapped with fresh film for 3 days. Dark brown necrotic lesions were observed on all inoculated branches 22 days post-inoculation. The controls had no symptoms. The pathogen was reisolated from all the inoculated branches, fulfilling Koch\'s postulates. To our knowledge, this is the first report of F. solani causing twig canker on English walnut in Xinjiang, China. Twig canker disease often cause a large number of branches to dry out and die. If the disease control and prevention is neglected, the productivity of the English walnut will be seriously affected in the cultivation area. Our finding will provide valuable information for prevention and management of twig canker on English walnut.
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  • 文章类型: Journal Article
    2020年8月,在核桃果园的核桃果和西格拉塔果实上观察到炭疽病病变,在宜军(陕西省)和南华(云南省)县,中国。核桃果实的症状首先表现为小的坏死点,迅速扩大为亚圆形或不规则凹陷的黑色病变(图1a,b).60个患病的核桃果实(30个J.regia和J.sigilata的果实,分别)从六个果园中随机抽样(每个果园10-15公顷,在每个县选择了三个果园),患有严重的炭疽病(果园中水果炭疽病的发病率超过60%。)在两个县。如Cai等人所述,从患病的果实中获得了26种单孢子分离物。(2009)。七天后,分离株形成灰色至乳白色菌落,菌落上表面有丰富的气生菌丝,PDA背面的乳白色到浅橄榄色(图。1c)。分生孢子细胞是透明的,光滑的壁,和圆柱形到棒形(图。1d).分生孢子是光滑的,无菌,圆柱形到梭形,两端锐角或一端圆形,一端略锐角(图。1e),尺寸范围为15.5-24.3×4.9-8.1µm(n=30)。Pressoria为棕色至中等棕色,棒形到椭圆形,边缘整个或起伏(图。1f),大小在8.0-27.6×4.7-13.7μm之间(n=30)。26个分离株的形态特征与杂病菌的形态特征相似(Damm等人。2012).随机选择六个代表性分离株(每个省三个分离株)用于分子分析。核糖体内部转录间隔区(ITS)(White等人。1990),β-微管蛋白(TUB2)(GlassandDonaldson1995),甘油醛-3-磷酸脱氢酶(GAPDH)(Templeton等人1992)和几丁质合酶1(CHS-1)(Carbone和Kohn1999)基因被扩增并测序。26个分离株中的6个的序列提交给GenBank(登录号:ITS:MT799938-MT799943,TUB:MT816321-MT816326,GAPDH:MT816327-MT816332,CHS-1:MT816333-816338)。多位点系统发育分析显示,六个分离株与炭疽病分离株CBS133.44和CBS130251聚集在一起,自举支持值为100%(图2)。两个代表性分离株(CFCC54247和CFCC54244)的致病性是使用健康水果进行测试的。香陵\“和\”J.西吉拉塔简历。杨碧“品种。用无菌针头刺穿核桃果皮,将40个灭菌水果(20个水果接种CFCC54247,20个水果接种CFCC54244)炸伤,并用10μl分生孢子悬浮液(106分生孢子/ml)从25℃的PDA上生长的7天大菌落接种在伤口部位。用无菌水接种20个受伤的果实作为对照。将接种的果实和对照果实在容器中在25℃下以12/12h的光/暗循环孵育。该实验重复三次。炭疽病症状(图。1g-h)在12天后在所有接种的水果中观察到,而对照组没有症状。从接种的病果中分离的真菌表现出与本研究中获得的分离株相同的形态和分子特征,确认科赫的假设。据我们所知,这是中国两种核桃引起炭疽病的首例报告。该结果将有助于为进一步研究该病的控制提供依据。
    In August 2020, anthracnose lesions were observed on fruits of Juglans regia and J. sigillata in walnut orchards, in Yijun (Shaanxi Province) and Nanhua (Yunnan Province) counties, China. Symptoms on walnut fruits first appeared as small necrotic spots that rapidly enlarged into subcircular or irregular sunken black lesions (Fig. 1a, b). Sixty diseased walnut fruits (30 fruits of J. regia and J. sigillata, respectively) were randomly sampled from six orchards (10-15 ha each orchard, three orchards were selected in each county) with severe anthracnose (incidence rate of fruit anthracnose is over 60% in the orchard.) in two counties. Twenty-six single spore isolates were obtained from diseased fruits as described by Cai et al. (2009). After seven days, isolates formed grey to milky white colony with abundant aerial hyphae on the upper surface of colony, and milky white to light olive on the back of PDA (Fig. 1c). Conidiogenous cells were hyaline, smooth-walled, and cylindrical to clavate (Fig. 1d). Conidia were smooth-walled, aseptate, cylindrical to fusiform, with both ends acute or one end round and one end slightly acute (Fig. 1e), and ranged in size from 15.5-24.3×4.9-8.1 µm (n=30). Appressoria were brown to medium brown, clavate to elliptical, with the edge entire or undulate (Fig. 1f), and ranged in size from 8.0-27.6×4.7-13.7μm (n=30). The morphological characteristics of 26 isolates were similar to those of the species complex Colletotrichum acutatum (Damm et al. 2012). Six representative isolates were randomly selected (three isolates for each province) for molecular analysis. The ribosomal internal transcribed spacers (ITS) (White et al. 1990), beta-tubulin (TUB2) (Glass and Donaldson 1995), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) (Templeton et al.1992) and chitin synthase 1 (CHS-1) (Carbone and Kohn 1999) genes were amplified and sequenced. Sequences of 6 of 26 isolates were submitted to GenBank (Accession Nos: ITS: MT799938-MT799943, TUB: MT816321-MT816326, GAPDH:MT816327-MT816332, CHS-1: MT816333-816338). Multi-locus phylogenetic analyses revealed that six isolates clustered together with Colletotrichum godetiae ex-type culture isolates CBS133.44 and CBS130251, and the bootstrap support value was 100% (Fig.2). The pathogenicity of two representative isolates (CFCC54247 and CFCC54244) was tested using healthy fruits of the \" J. regia cv. Xiangling\" and \" J. sigillata cv. Yangbi\" varieties. Forty sterilized fruits (20 fruits were inoculated with CFCC54247, and 20 fruits with CFCC54244) were wounded by puncturing with a sterile needle through walnut pericarp and inoculated in the wound site with 10 μl of conidial suspension (106 conidia/ml) from seven day old colonies grown on PDA at 25℃. Twenty wounded fruits were inoculated with sterile water as control. Inoculated and control fruits were incubated in containers at 25℃ in a 12/12h light/dark cycle. The experiment was repeated three times. Anthracnose symptoms (Fig. 1g-h) were observed in all inoculated fruits after 12 days, whereas controls showed no symptoms. Fungal isolates from inoculated diseased fruits showed the same morphological and molecular characteristics as the isolates obtained in this study, confirming Koch\'s postulates. To our knowledge, this is the first report of C. godetiae causing anthracnose on the two walnut species in China. The result will be helpful for providing a basis for further research on the control of the disease.
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  • 文章类型: Journal Article
    山核桃(CaryaillinoinensisK.Koch)是江苏省重要且广泛种植的坚果树种,中国(莫等人。2018)。2020年7月,在裕廊山核桃上经常观察到叶斑病症状,江苏省(东经119°15\'36\“,32°1\'6\“N)。在来自三个苗圃的150棵成熟山核桃中,病害发生率为40%至65%。疾病严重程度指数(DSI,Jiang等人。2019年)达到58.4。症状开始时,叶子上散布着小的棕色斑点,逐渐扩大到大,圆形到不规则的黑色和棕色坏死病变。在严重的情况下,在单叶的大部分上形成的病变,最后枯叶从树上掉下来。从病变边缘分离出三个单分生孢子分离株(Chen2346,Chen2347,Chen2348),并在马铃薯葡萄糖琼脂(PDA)培养基上培养。PDA上的殖民地是白色和棉质的,后来变成浅灰色,具有丰富的生殖结构。孢子囊是无盐的,透明玻璃,无分支,顶端扩张形成锁骨状囊泡,产生了孢子囊。孢子囊呈球状至椭圆形,棕色到深棕色,长度为103至128µm,宽度为88至114微米(n=30)。孢子虫是棕色的,椭圆形到卵圆形,有纵向条纹,测量为13.9至18.8×7.9至10.8μm(n=60)。这些分离株的形态特征与葫芦的描述一致(Kirk1984)。提取了这三个单分生孢子分离株的基因组DNA,使用引物对ITS1/ITS4扩增rDNA的内部转录间隔区(ITS)和大亚基(LSU)的部分序列(Whiteetal。1990)和LR0R/LR7(Vilgalys和Hester1990),分别。共有序列(GenBank登录号:使用BLASTn比对ITS的OP315248至OP315250和LSU的OP315251至OP315253),并与GenBank中发现的葫芦C.的序列显示出100%的同一性(登录号。:MF942131用于ITS和ON025788用于LSU)。为了进一步确认身份,使用MEGA(v.7.0)和MrBayes(v.3.1.2)软件进行系统发育分析,使用最大似然和贝叶斯推断方法,分别。多基因系统发育表明,本研究中的三个分离株,还有,C.葫芦标本,群集为强烈支持的单系组(99个引导值;0.95个后验概率)。根据形态学和分子数据,分离株被鉴定为葫芦。为了确认致病性,健康的山核桃幼苗(2岁)分别接种菌丝体块(3×3毫米),从每片叶子的静脉上的菌落边缘切除。用非定殖的PDA块处理的幼苗用作对照。将接种的幼苗在25±1°C和12小时光周期下保持在无菌塑料盒中,其中装有湿润的滤纸片。对于每种处理,测试每种分离物15片叶。该实验重复两次。接种三天后,类似于现场出现的症状,而对照叶保持无症状。随后,从病变中重新分离出葫芦杆菌,并进行形态学鉴定,确认科赫的假设。据我们所知,这是在中国首次报道了葫芦叶斑病。本研究为进一步研究生物学提供了基础,流行病学,和这种疾病的管理。
    Pecan (Carya illinoinensis K. Koch) is an important and widely planted nut tree species in Jiangsu Province, China (Mo et al. 2018). In July 2020, leaf spot symptoms were frequently observed on pecan in Jurong, Jiangsu Province (119°15\'36\"E, 32°1\'6\"N). Disease incidences ranged from 40 to 65% among 150 mature pecan trees from three nurseries. The disease severity index (DSI, Jiang et al. 2019) reached 58.4. Symptoms began as small brown spots scattered on leaves that gradually expanded to large, circular to irregular black and brown necrotic lesions. In severe cases, lesions developed on large portions of a single leaf, and eventually the dead leaves fell from the trees. Three monoconidial isolates (Chen2346, Chen2347, Chen2348) were isolated from lesion margins and cultured on potato dextrose agar (PDA) medium. Colonies on PDA were white and cottony, later becoming light gray with abundant reproductive structures. Sporangiophores were aseptate, hyaline, unbranched, and apically dilated to form a clavate vesicle, which produced sporangia. Sporangia were globular to ellipsoid, brown to dark brown, 103 to 128 µm in length, and 88 to 114 µm in width (n = 30). Sporangiola were brown, ellipsoid to ovoid, with longitudinal striae, and measured 13.9 to 18.8 × 7.9 to 10.8 μm (n = 60). The morphological characteristics of these isolates agreed with descriptions of Choanephora cucurbitarum (Kirk 1984). Genomic DNA of these three monoconidial isolates was extracted, and the partial sequences of the internal transcribed spacer (ITS) and large subunit (LSU) of rDNA were amplified using primer pairs ITS1/ITS4 (White et al. 1990) and LR0R/LR7 (Vilgalys and Hester 1990), respectively. The consensus sequences (GenBank accession nos.: OP315248 to OP315250 for ITS and OP315251 to OP315253 for LSU) were aligned using BLASTn and showed100% identity with the sequences from C. cucurbitarum found in GenBank (accession nos.: MF942131 for ITS and ON025788 for LSU). To further confirm the identity, a phylogenetic analysis was performed with MEGA (v.7.0) and MrBayes (v.3.1.2) software, using the maximum likelihood and Bayesian inference methods, respectively. The multigene phylogeny revealed that the three isolates in this study, as well as, C. cucurbitarum specimen, clustered as a strongly supported monophyletic group (99 bootstrap value; 0.95 posterior probabilities). Based on the morphological and molecular data, the isolates were identified as C. cucurbitarum. To confirm pathogenicity, healthy pecan seedlings (2 years old) were each inoculated with a mycelial block (3 × 3 mm) excised from the margin of a colony on the vein of each leaf. Seedlings treated with non-colonized PDA blocks were used as controls. The inoculated seedlings were maintained in sterile plastic boxes with moistened sheets of filter paper at 25 ± 1°C and a 12-h photoperiod. Fifteen leaves per isolate were tested for each treatment. The experiment was repeated twice. Three days after inoculation, symptoms similar to those in the field appeared, whereas the control leaves remained symptomless. Subsequently, C. cucurbitarum was reisolated from the lesions and morphologically identified, confirming Koch\'s postulates. To the best of our knowledge, this is the first report of C. cucurbitarum causing leaf spot on C. illinoinensis in China. This study provides the foundation to further investigate the biology, epidemiology, and management of this disease.
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    铁核桃(JuglanssigillataDode)是中国特有的温带落叶乔木。它主要分布在中国西南部,并因其木材和坚果而受到重视(Feng等人。2018)。2020年9月,在崇州市占地2公顷的果园中观察到J.sigilata的溃疡症状,四川省(北纬31°5\'25″,105°27\'36″E,365米高度)。在接受调查的50种植物中,有20%的植物表现出溃疡症状。感染的躯干显示坏死病变,伴有黑色比尼迪病,在严重的情况下导致树枝坏死和整个植物死亡(图。1).收集不同病种植物的6个标本进行病原分离和形态学观察。根据Chomnunti(Chomnunti等人)描述的方法,从马铃薯-葡萄糖琼脂(PDA)培养基上的单个分生孢子中获得纯培养物。2014).菌落生长迅速,5天后达到3厘米。气生菌丝丰富,最初是白色,然后是灰色。在宿主上的分生孢子测量为160-280µm×140-190µm(平均:220×165µm,n=20),基质,单身,深棕色到黑色,沉浸,成熟的时候就会发臭。Pycnidial墙32-58µm宽,由5-7层棕色至深棕色细胞组成。分生孢子是透明的,椭圆形,顶端和基部圆形,中间最宽,厚壁,和单细胞,尺寸为21.5-31µm×11.5-15.7µm(平均:27×13.5µm,n=50)。形态特征符合Lasiodiplodia伪TheobromaeA.J.L.Phillips的描述,A.Alves&Crous(Aives等人。2008).内部转录间隔区(ITS),18S小亚基rRNA(SSU),28S大亚基rDNA(LSU),翻译延伸因子1-α(tef1-α),和β-微管蛋白(tub2)通过聚合酶链反应扩增,并用引物ITS1/ITS4,NS1/NS4,LR0R/LR5,EF1-728F/EF1-986R和Bt2a/Bt2b进行测序,分别(Li等人。2018)。代表性分离株(SICAUCC22-0079)的序列保藏于NCBI,登录号ON090365(ITS),ON090406(SSU),ON090418(LSU),ON112377(tef1-α),和ON112378(tub2),分别。核苷酸爆炸显示,所有分析和NCBI提交的分离株与假可可乳杆菌(CBS116459;整型)(登录号EF622077,EU673199,EU673256,EF622057,EU673111)具有100%的相似性。基于组合数据集的系统发育分析显示,在具有假性乳杆菌复合物的进化枝中,100%的Bootstrap支持值(图。2).基于形态学和分子分析,真菌病原体被鉴定为假性乳杆菌。为了执行科赫的假设,用10μL孢子悬浮液(105分生孢子/mL)通过无菌针将四株2岁的健康植物接种到受伤部位。作为控制,用无菌蒸馏水处理四株健康植物。将接种和未处理的植物置于25°C、相对湿度>90%和12小时光周期的生长室中。接种植物15-20天后出现树干溃疡症状,病原体被重新分离,对照没有症状,确认科赫的假设。假可可乳杆菌广泛分布于世界各地的各种植物中,通常作为与阻尼相关的病原体,wilt,死回,根腐病,项圈腐烂,女巫\'扫帚,或水果腐烂(赵等人。2010).据我们所知,这是在中国首次报道了由假可可乳杆菌引起的J.sigilata树干溃疡。由假拟人乳杆菌引起的树干溃疡正在成为核桃生产的潜在威胁,并应采取一些必要的综合管理措施。
    Iron walnut (Juglans sigillata Dode) is a temperate deciduous tree indigenous to China. It is mainly distributed in southwestern China, and valued for its wood and nuts (Feng et al. 2018). In September 2020, symptoms of canker on J. sigillata were observed in an orchard measuring 2 hectares located in Chongzhou City, Sichuan Province (31°5\' 25″N, 105°27\'36″E, 365 m altitude). Twenty percent of plants showed canker symptoms during the 50 surveyed plants. The infected trunk showed necrotic lesions with black pycnidia, that led to necrosis of branches and death of the whole plant in severe cases (Fig. 1). Six specimens from different diseased plants were collected for pathogen isolation and morphological observation. Pure cultures were obtained from single conidium on potato-dextrose agar (PDA) media according to the method described by Chomnunti (Chomnunti et al. 2014). Colonies grew fast and reached 3 cm after 5 days. The aerial mycelium was abundant, which was initially white and then grayish. Conidiomata on the host were measured 160-280 µm × 140-190 µm (average: 220 × 165 µm, n = 20), stromatic, uniloculate, dark brown to black, immersed, and erumpent when mature. Pycnidial walls 32-58 µm wide, were composed of 5-7 layers of brown to dark brown cells. Conidia were hyaline, and ellipsoidal with rounded apex and base, widest at the middle, thick-walled, and unicellular, with a size 21.5-31 µm × 11.5-15.7 µm (average: 27 × 13.5 µm, n = 50). Morphological characteristics fit the description of Lasiodiplodia pseudotheobromae A.J.L. Phillips, A. Alves & Crous (Aives et al. 2008). The internal transcribed spacers (ITS), 18S small subunit rRNA (SSU), 28S large subunit rDNA (LSU), translation elongation factor 1-alpha (tef1-α), and beta-tubulin (tub2) were amplified by polymerase chain reaction and sequenced with primers ITS1/ITS4, NS1/NS4, LR0R/LR5, EF1-728F/EF1-986R and Bt2a/Bt2b, respectively (Li et al. 2018). The sequences of the representative isolate (SICAUCC 22-0079) were deposited in NCBI with accession numbers ON090365 (ITS), ON090406 (SSU), ON090418 (LSU), ON112377 (tef1-α), and ON112378 (tub2), respectively. Nucleotide blast showed 100% similarity of all the analyzed and NCBI submitted isolates with L. pseudotheobromae (CBS116459; holotype) (accession numbers EF622077, EU673199, EU673256, EF622057, EU673111). Phylogenetic analyses based on a combined dataset showed 100% bootstrap support values in a clade with L. pseudotheobromae complexes (Fig. 2). Based on morphological and molecular analyses, the fungal pathogen was identified as L. pseudotheobromae. To conduct Koch\'s postulates, four 2-year-old healthy plants of J. sigillata were inoculated with 10 μL spore suspension (105 conidia/mL) onto the wounded sites via sterile pin. As control, four healthy plants were treated with sterile distilled water. The inoculated and untreated plants were placed in a growth chamber at 25°C with relative humidity >90% and 12-h photoperiod. Trunk canker symptoms appeared on inoculated plants after 15-20 days, and the pathogen was re-isolated and the controls were symptomless, confirming Koch\'s postulates. L. pseudotheobromae is widely distributed in various plants all over the world, usually as a pathogen associated with damping-off, wilt, die-back, root rot, collar rot, witches\' brooms, or fruit rots (Zhao et al. 2010). To our knowledge, this is the first report of trunk canker on J. sigillata caused by L. pseudotheobromae in China. Trunk canker caused by L. pseudotheobromae is becoming a potential threat to walnut production, and some necessary measures for integrated management should be made.
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    JuglanssigilataDode,一种原产于中国西南地区的特有核桃,在四川省主要用作坚果(Jin等人。,2019)。2021年5月,在绵阳市10公顷的果园中观察到分支枯萎病的症状,四川省(北纬31°5\'25″,105°27\'36″E,海拔365m)。大约40%的植物在由20棵核桃树组成的四边形中患病,每棵受影响的树上有20%的树枝死亡。最初,出现浅棕色斑点;然后,斑点扩大到围绕整个树枝;最后,树枝从棕色变成红棕色并死亡。从不同的树上随机取样四个有症状的分支。接下来,使用单分生孢子分离从每个分支的尖刺中获得了四个真菌分离株(Chomnunti等人。2014年),并在马铃薯葡萄糖琼脂(PDA)上培养。将培养皿置于培养箱中并在25°C下在12小时光周期下培养。菌落最初为白色,有稀薄的气生菌丝体,逐渐变成深灰色,边缘不规则。分生孢子是囊状,黑色和分散,直径为0.3-0.7毫米。分生孢子是窄圆柱形的,在底部简单或分支,30-43×3-8µm(x=36.5×5.5µm,n=40)。分生孢子细胞是具有明显环状的annellic。分生孢子是单细胞的,成熟时呈棕色,狭椭圆体,基部有凝胶状鞘和截短疤痕,17-32×7-12µm(x=27×9µm,n=40)。提取代表性分离株SICAUCC22-0064的基因组DNA,和内部转录间隔区(ITS),鸟嘌呤核苷酸结合蛋白亚基β基因(ms204),翻译延伸因子1-α(tef1-α),通过聚合酶链反应扩增β-微管蛋白(tub2)的部分序列,并用引物V9G/LR5测序(deHoog&vandenEnde1998),MS-E1F1/MS-E5R1(Walker等.2012),EF1-728F(Carbone&Kohn1999)/TEF1LLErev(Jaklitsch等人2005),和T1/BtHV2r(Voglmayr等人2017),分别。ITS的序列,ms204,tef1-α,和tub2分别存放在NCBI,登录号为ON000068,ON112376,ON112374和ON112375。考虑到JuglconisappreculataVoglmayr和Jaklitsch的前型菌株(D96)中ms204和tub2的序列缺失,分离物D140用于核苷酸胚细胞。结果显示99.68%,100%,100%,和100%的ITS身份,ms204,tef1-α,和带有D140的tub2(登录号KY427138、KY427157、KY427207、KY427226)。基于组合数据集的系统发育分析显示100%自举与阑尾J.形态与阑尾J的无性阶段一致(Voglmayr等人。,2017)。为了验证科赫的假设,在每株植物中用分生孢子悬浮液(1×105分生孢子/mL)喷洒被针刺伤的五个分支,在健康的2岁盆栽植物上进行了三次重复。用无菌蒸馏水喷洒相同数量的分支作为对照。将植物置于25°C、90%相对湿度和12小时荧光光/暗状态下的温室中。五周后,所有接种的分支都显示出与田间观察到的相似的棕色坏死,对照组没有出现症状。从坏死病变中重新分离出病原体,并通过形态学和系统发育进行鉴定。J.阑尾在奥地利的Juglansnigra和J.regia报告,法国,西班牙和希腊(Farr&Rossman2022)。本文是中国首例阑尾J.U.S.引起的核桃枝枯病的报道。这一结果可以加深对核桃病害的认识,为进一步管理奠定基础。
    Juglans sigillata Dode, an endemic walnut species native in southwest China, is mainly used as nuts in Sichuan Province (Jin et al., 2019). In May 2021, symptoms of branches blight were observed in an orchard measuring 10 hectares located in Mianyang City, Sichuan Province (31°5\' 25″N, 105°27\'36″E, 365 m above sea level). About 40% of plants were diseased in the quadrat consisting of twenty walnut trees, and 20% of branches were dead on each affected tree. Initially, light brown spots appeared; then, the spots expanded to surround the whole branches; finally, the branches changed from brown to reddish-brown and died. Four symptomatic branches were sampled randomly from different trees. Next, four fungal isolates were obtained from the acervuli of each branch using the single-conidium isolation (Chomnunti et al. 2014) and cultured on potato dextrose agar (PDA). The Petri dishes were placed in an incubator and cultured at 25 °C under a 12-h photoperiod. Colonies were initially white with thin aerial mycelia and gradually turned dark grey with irregular margins. Conidiomata were acervular, black and scattered, with a diameter of 0.3 - 0.7 mm. Conidiophores were narrowly cylindrical, simple or branched at the base, 30 - 43 × 3 - 8 µm (x = 36.5 × 5.5 µm, n = 40). Conidiogenous cells were annellidic with distinct annellations. Conidia were unicellular, brown when mature, narrowly ellipsoid with gelatinous sheaths and truncate scars at the base, 17 - 32 × 7 - 12 µm (x = 27 × 9 µm, n = 40). The genomic DNA of a representative isolate SICAUCC 22-0064 was extracted, and the internal transcribed spacer (ITS) region, guanine nucleotide-binding protein subunit beta gene (ms204), translation elongation factor 1-alpha (tef1-α), and partial sequences of β-tubulin (tub2) were amplified by polymerase chain reaction and sequenced with primers V9G/LR5 (de Hoog & van den Ende 1998), MS-E1F1/MS-E5R1 (Walker et al. 2012), EF1-728F (Carbone & Kohn 1999)/TEF1LLErev (Jaklitsch et al. 2005), and T1/BtHV2r (Voglmayr et al. 2017), respectively. The sequences of ITS, ms204, tef1-α, and tub2 were deposited in NCBI with accession numbers ON000068, ON112376, ON112374, and ON112375, respectively. With the consideration of the sequence lack of ms204 and tub2 in the ex-type strain (D96) of Juglanconis appendiculata Voglmayr & Jaklitsch, the isolate D140 was used for nucleotide blast. The results showed 99.68%, 100%, 100%, and 100% identities of ITS, ms204, tef1-α, and tub2 with D140 (accession numbers KY427138, KY427157, KY427207, KY427226). Phylogenetic analysis based on a combined dataset showed 100% bootstrap with J. appendiculata, and the morphology was consistent with the asexual stage of J. appendiculata (Voglmayr et al., 2017). To verify Koch\'s postulates, five branches wounded by pin-prick were sprayed with conidial suspension (1 × 105 conidia/mL) in each plant, and three repetitions were performed on healthy 2-year-old potted plants. The same number of branches were sprayed with sterile distilled water as controls. The plants were placed in a greenhouse at 25 ℃ under 90% relative humidity and a 12-h fluorescent light/dark regime. After five weeks, all the inoculated branches showed brown necrosis similar to that observed in the field, and no symptoms occurred on the controls. The pathogens were re-isolated from the necrotic lesions and identified by morphology and phylogeny. J. appendiculata has been reported on Juglans nigra and J. regia in Austria, France, Spain and Greece (Farr & Rossman 2022). This paper is the first report of branch blight on Juglans sigillata caused by J. appendiculata in China. This result may develop the understanding of walnut diseases and lay a foundation for further management.
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    JuglansregiaL.是中国坚果和木材的主要栽培核桃品种之一(Pollegioni等人。2012).2020年6月,在崇州市一果园(30°33\'34″N,103°38\'35英寸)。在30公顷的果园里,发病率约为50%。总共取样了15株植物,其40%的分支受到这种疾病的影响。首先,出现棕色和不规则斑点,然后斑点逐渐扩大并包围了树枝,最终杀死了树枝.收集了来自不同树木的患病树枝的五个样品,并使用单子囊孢子分离法从五个样品中的每个样品中获得了单个真菌分离物(Chomnunti等人。2014).马铃薯葡萄糖琼脂(PDA)上的五个分离株的菌落相同,最初在顶部出现白色,随着年龄的增长变成浅棕色到深棕色。在主机上,胚乳是黑色的,球状至亚球状,短乳头状,卵形,260-410×210-320µm(x=335×265µm,n=20)。Asci有8个孢子,bitunicate,圆柱形,短花梗,55-78×8-12µm(x=67.5×10µm,n=40)。子囊孢子是1间隔的,梭形到椭圆形,略微弯曲,波形,12-17×3-5微米(x=14.5×4微米,n=40)。这些性形态特征与PalmiascomaqujingensePhook一致。&K.D.Hyde(Monkai等人。2021)。17天后在培养箱中的PDA上形成无性形态(25℃,90%相对湿度,12小时光周期)。分生孢子是黑色的,球状至亚球状,220-300×240-380µm(x=270×310µm,n=20)。分生孢子为椭圆形,光滑和光滑的壁,3-7×2-4μm(x=4.9×3μm,n=50)。提取代表性分离株SICAUCC21-0013的基因组DNA,和内部转录间隔区(ITS),大亚基rDNA(LSU)区,小亚基rDNA(SSU)区,和RNA聚合酶II(rpb2)基因的最大亚基被扩增并用引物ITS5/ITS4测序(Whiteetal。1990),LR0R/LR5(Rehner等人1994),NS1/NS4(White等人1990),和fRPB2-5F/fRPB2-7cR(Liu等人1999),分别。序列保藏于NCBI,登录号分别为MZ983549、MZ959419、MZ951112和MZ818772,显示100%,100%,99.14%,和99.59%的身份与P.qujingenseKUMCC19-0201(整型)(登录号分别为MT477185、MT477186、MT477183、MT495782)。基于串联数据集的系统发育分析(最大似然)显示,P.qujingense具有93%的自举支持值。为了验证科赫的假设,通过针刺接种,用来自4周龄培养物的分生孢子悬浮液(106分生孢子/ml)接种了来自三个1岁幼苗的9个健康分支(Desai等人。2019),用无菌水接种相同数量的幼苗和树枝作为对照。将植物置于25°C和90%RH的温室中,在12小时的荧光/黑暗状态下。28天后,在接种P.qujingense的树枝上形成褐色斑点,与在田间观察到的相似,而对照组则无症状。从病变中重新分离出病原体,并通过形态学和系统发育进行鉴定。据我们所知,这是世界上首例P.qujingense引起J.regia分支疫病的报告。这种疾病可能会影响J.regia的生长和产量,并应采取控制措施。
    Juglans regia L. is one of the major cultivated walnut species in China for nuts and wood (Pollegioni et al. 2012). In June 2020, branches with blight symptoms were observed in an orchard at Chongzhou City (30°33\'34″N, 103°38\'35″E). In an orchard of 30 hectares, disease incidence was around 50%. A total of 15 plants were sampled and 40% of their branches were affected by this disease. Firstly, brown and irregular spots appeared, then the spots gradually expanded and encircled the branch, which eventually killed the branch. Five samples of diseased branches from different trees were collected and a single fungal isolate was obtained from each of the five samples using the single ascospore isolation (Chomnunti et al. 2014). Colonies of the five isolates on potato dextrose agar (PDA) were identical that initially appeared white on the top, becoming light to dark brown with age. On the host, ascostroma were black, globose to subglobose, short-papillate, ostiolate, 260 - 410 × 210 - 320 µm (x = 335 × 265 µm, n = 20). Asci were 8-spored, bitunicate, cylindrical, short pedicellate, 55 - 78 × 8 - 12 µm (x = 67.5 × 10 µm, n = 40). Ascospores were 1-septate, fusiform to ellipsoidal, slightly curved, guttulate, 12 - 17 × 3 - 5 µm (x = 14.5 × 4 µm, n = 40). These sexual morphological characteristics are consistent with the Palmiascoma qujingense Phook. & K.D. Hyde (Monkai et al. 2021). Asexual morphs were formed on PDA in incubator after 17 days (25℃, 90% relative humidity, 12-h photoperiod). Conidiomata were black, globose to subglobose, 220 - 300 × 240 - 380 µm (x = 270 × 310 µm, n = 20). Conidia were oblong to ellipsoidal, aseptate and smooth-walled, 3 - 7 × 2 - 4 μm (x = 4.9 × 3 μm, n = 50). The genomic DNA of a representative isolate SICAUCC 21-0013 was extracted, and the internal transcribed spacers (ITS) region, large subunit rDNA (LSU) region, small subunit rDNA (SSU) region, and the largest subunit of RNA polymerase II (rpb2) gene were amplified and sequenced with primers ITS5/ITS4 (White et al. 1990), LR0R/LR5 (Rehner et al. 1994), NS1/NS4 (White et al. 1990), and fRPB2-5F/fRPB2-7cR (Liu et al. 1999), respectively. The sequences were deposited in NCBI with accession numbers MZ983549, MZ959419, MZ951112, and MZ818772, respectively, which showed 100%, 100%, 99.14%, and 99.59% identities with P. qujingense KUMCC 19-0201 (holotype) (accession numbers MT477185, MT477186, MT477183, MT495782respectively). Phylogenetic analysis (maximum likelihood) based on a concatenated dataset showed 93% bootstrap support values with P. qujingense. To verify Koch\'s postulates, 9 healthy branches from three 1-year-old seedlings were inoculated with conidial suspension (106 conidia/ml) from 4-week-old cultures via pin-prick inoculation (Desai et al. 2019), and the same number of seedlings and branches were inoculated with sterile water as controls. Plants were placed in a greenhouse at 25℃ and 90% RH on a 12-h fluorescent light/dark regime. After 28 days, brown spots were formed on P. qujingense-inoculated branches and similar to those observed in the field, while the controls remained asymptomatic. The pathogen was re-isolated from the lesions and identified by morphology and phylogeny. To our knowledge, this is the first report of P. qujingense causing branch blight on J. regia in the world. This disease potentially impacts the growth and yield of J. regia, and control measures should be made.
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    使用营养干预措施来管理糖尿病是旨在降低全球患病率的有效策略之一。正在上升。杏仁是食用最多的树坚果,众所周知它们富含蛋白质,单不饱和脂肪酸,必需矿物质,和膳食纤维。因此,这篇综述的目的是评估杏仁对肠道微生物群的影响,糖代谢,2型糖尿病患者的炎症参数。
    方法:根据系统评价和荟萃分析(PRISMA)的首选报告项目进行系统评价和荟萃分析。EBSCOhost,其中包括健康科学研究数据库;谷歌学者;EMBASE;并根据人口搜索文章的参考列表,干预,control,结果,和研究(PICOS)框架。从数据库开始到2021年8月1日,根据医学主题词(MesH)和同义词进行了搜索。使用ReviewManager(RevMan)5.3软件进行荟萃分析。
    结果:系统评价中纳入了9项随机研究,其中8项用于荟萃分析。结果表明,基于杏仁的饮食在促进产生短链脂肪酸(SCFA)的肠道微生物群的生长方面具有显着的作用。此外,荟萃分析显示,以杏仁为基础的饮食可显著降低(p<0.05)2型糖尿病患者的糖化血红蛋白(HbA1c)水平和体重指数(BMI).然而,还发现杏仁对空腹血糖的影响不显著(p>0.05),餐后2小时血糖,炎症标志物(C反应蛋白和肿瘤坏死因子α,TNF-α),胰高血糖素样肽-1(GLP-1),胰岛素抵抗的稳态模型评估(HOMA-IR),和空腹胰岛素。在这篇综述中观察到的与降低HbA1c和BMI有关的结果的生物学机制可能是基于杏仁的营养成分和生物学效应,包括高纤维含量和低血糖指数。
    结论:本系统综述和荟萃分析的结果表明,与对照组相比,以杏仁为基础的饮食可有效促进2型糖尿病患者产生短链脂肪酸的细菌,降低糖化血红蛋白和体重指数。然而,杏仁对空腹血糖的影响不显著(p>0.05),餐后2小时血糖,炎症标志物(C反应蛋白和TNF-α),GLP-1,HOMA-IR,和空腹胰岛素。
    The use of nutritional interventions for managing diabetes is one of the effective strategies aimed at reducing the global prevalence of the condition, which is on the rise. Almonds are the most consumed tree nut and they are known to be rich sources of protein, monounsaturated fatty acids, essential minerals, and dietary fibre. Therefore, the aim of this review was to evaluate the effects of almonds on gut microbiota, glycometabolism, and inflammatory parameters in patients with type 2 diabetes.
    METHODS: This systematic review and meta-analysis was carried out according to the preferred reporting items for systematic review and meta-analysis (PRISMA). EBSCOhost, which encompasses the Health Sciences Research Databases; Google Scholar; EMBASE; and the reference lists of articles were searched based on population, intervention, control, outcome, and study (PICOS) framework. Searches were carried out from database inception until 1 August 2021 based on medical subject headings (MesH) and synonyms. The meta-analysis was carried out with the Review Manager (RevMan) 5.3 software.
    RESULTS: Nine randomised studies were included in the systematic review and eight were used for the meta-analysis. The results would suggest that almond-based diets have significant effects in promoting the growth of short-chain fatty acid (SCFA)-producing gut microbiota. Furthermore, the meta-analysis showed that almond-based diets were effective in significantly lowering (p < 0.05) glycated haemoglobin (HbA1c) levels and body mass index (BMI) in patients with type 2 diabetes. However, it was also found that the effects of almonds were not significant (p > 0.05) in relation to fasting blood glucose, 2 h postprandial blood glucose, inflammatory markers (C-reactive protein and Tumour necrosis factor α, TNF-α), glucagon-like peptide-1 (GLP-1), homeostatic model assessment of insulin resistance (HOMA-IR), and fasting insulin. The biological mechanisms responsible for the outcomes observed in this review in relation to reduction in HbA1c and BMI may be based on the nutrient composition of almonds and the biological effects, including the high fibre content and the low glycaemic index profile.
    CONCLUSIONS: The findings of this systematic review and meta-analysis have shown that almond-based diets may be effective in promoting short-chain fatty acid-producing bacteria and lowering glycated haemoglobin and body mass index in patients with type 2 diabetes compared with control. However, the effects of almonds were not significant (p > 0.05) with respect to fasting blood glucose, 2 h postprandial blood glucose, inflammatory markers (C-reactive protein and TNF-α), GLP-1, HOMA-IR, and fasting insulin.
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  • 文章类型: Journal Article
    澳洲坚果(Macadamiaternifolia)最早于1910年从澳大利亚传入中国,主要栽培区为云南和广西。它既可以用作水果,也可以用作治疗药物,具有很高的经济价值。在2021年3月,观察到M.ternifolia展示了女巫\'-扫帚,德宏州的叶片黄化和丛丛芽,云南省,中国。受感染植物的顶芽被抑制,侧芽被刺激提前发芽为小枝。它被命名为M.ternifolia女巫\'-扫帚病,并在芒市的城市和农村地区被发现,梁河,迎江,芒东,龙川和龙陵市县。在所调查的七个地区,超过40%的植物被感染。将有症状和无症状植物的侧茎切成小块。通过固定处理组织,脱水和喷金。并在扫描电子显微镜(HitachiS-3000N)下观察组织(Pathan等人。2010).在有症状植物的韧皮部筛细胞中发现了近球形体。从七个地区收集有症状和无症状的植物,ddH2O用作阴性对照,和杜氏粘胶巫术\'-扫帚病植物被用作阳性对照。使用CTAB方法从0.1g组织中进行总植物DNA提取(Porebski等人。1997),并在西南林业大学森林灾害预警与控制重点实验室的冰箱中储存在-20℃。巢式PCR用于用引物P1/P7和R16F2/R16R2扩增16SrRNA基因(Lee等人。1993;Schneider等人。1993).扩增了1.8kb和1.2kb的PCR扩增子(GenBank材料MW892818、MW892819、MW892820、MW892821)。使用引物对rp(I)F/rp(I)R的直接PCR(Lee等人。2003年),对核糖体蛋白(rp)基因具有特异性,产生了约1.2kb的扩增子(GenBank加入MZ442600,MZ442601,MZ442602,MZ442603)。来自21个样品的片段与阳性对照一致,确认植物质与疾病的关联。有趣的是,从4份无症状植物样品中扩增出>16SrRNA基因和rp基因,我们推测澳洲坚果存在潜伏感染和隐藏症状(Moslemkhani和Sadeghi2011)。对MTWB植原虫16SrRNA序列的BLAST分析表明,它与Tremalaevigata女巫\'-broom植原虫具有99%的相似性(GenBank登录号MG755412)。rp序列与\'柳柳四腺\'女巫\'-扫帚植原体具有99%的同一性(GenBank登录号KC117314)。用iPhyClassifier进行的分析表明,从MTWB植原虫的查询16SrDNA片段得出的虚拟RFLP模式与16SrI组的参考模式最相似,亚组B(OY-M,GenBank登录号AP006628),模式相似系数为0.99。植物质被鉴定为Ca。属16SrI-B亚组的星型植物相关菌株通过使用MEGA版本6.0,基于16SrRNA基因和rp基因序列构建了系统发育树(Tamura等人。2013)与邻居加入(NJ)方法和引导支持估计有1000个重复。结果表明,MTWB植物质分别在16SrI-B和rpI-B中形成亚分化。2013年,澳洲坚果显示出由Ca引起的叶片硬度叶状和芽增殖。阿特米萨的星型植物,古巴。令人担忧的是,澳洲坚果植原体与16SrI-F亚组密切相关,它与中国菌株显著不同(Pérez-López等人。2013).此外,在苗圃条件下,MTWB植物浆从受感染的植物嫁接传播到健康植物(Akhtar等人。2009;Ikten等人。2014).在巢式PCR检测中,嫁接植物的植原体呈阳性。值得注意的是,植物受到蚜虫的严重破坏,据推测,同翅目昆虫是通过吸吮植物汁液引起疾病的传播,因此,必须确定在中国传播MTWB的蚜虫,以控制三联草女巫\'-扫帚病。据我们所知,澳洲坚果是钙的新寄主植物。中国的星型植物。新出现的疾病是对澳洲坚果的威胁。
    Macadamia nut (Macadamia ternifolia) was first introduced into China from Australia in 1910, and the main cultivation areas were Yunnan and Guangxi. It can be used as both a fruit and a therapeutic drug, with high economic value. In March 2021, it was observed that the M. ternifolia was showing witches\'-broom, leaf yellowing and plexus bud in Dehong Prefecture, Yunnan Province, China. The terminal buds of infected plants were inhibited and the lateral buds are stimulated to germinate into twigs in advance. It was named the M. ternifolia witches\'-broom disease, and was found in urban and rural areas of Mangshi, Lianghe, Yingjiang, Mangdong, Longchuan and Longling cities and counties. More than 40% of the plants were infected on the seven areas surveyed. The lateral stems from symptomatic and asymptomatic plants were cut to small pieces. The tissues were treated by fixation, dehydration and spraying-gold. And the tissues were observed under a scanning electron microscope (Hitachi S-3000N) (Pathan et al. 2010). The nearly spherical bodies were found in the phloem sieve cells of symptomatic plants. Symptomatic and asymptomatic plants were collected from seven areas, ddH2O was used as the negative control, and Dodonaea viscose witches\'-broom disease plants were used as the positive control. The total plants\' DNA extraction was conducted from 0.1 g tissue using the CTAB method (Porebski et al. 1997), and were stored at -20 °C in a refrigerator in the Key Laboratory of Forest Disaster Warning and Control at the Southwest Forestry University. The nested PCR was employed to amplify the 16S rRNA gene with the primers P1/P7 and R16F2/R16R2 (Lee et al. 1993; Schneider et al. 1993). PCR amplicon of 1.8 kb and 1.2 kb were amplified (GenBank accessions MW892818, MW892819, MW892820, MW892821). The direct PCR with primer pairs rp(I)F/ rp(I)R (Lee et al. 2003) specific to the ribosomal protein (rp) gene yielded amplicons of approximately 1.2 kb (GenBank accessions MZ442600, MZ442601, MZ442602, MZ442603). The fragment from 21 samples was consistent with the positive control, confirming the association of phytoplasma with the disease. Interestingly, the phytoplasma/span>16S rRNA gene and rp gene was also amplified from the 4 samples of asymptomatic plant, we speculated that the latent infection and hidden symptoms existed in Macadamia nut (Moslemkhani and Sadeghi 2011). A BLAST analysis of the 16S rRNA sequences of MTWB phytoplasma showed that it has a 99% similarity with Trema laevigata witches\'-broom phytoplasma (GenBank accession MG755412). The rp sequence shared 99% identity with \'Salix tetradenia\' witches\'-broom phytoplasma (GenBank accession KC117314). An analysis with iPhyClassifier showed that the virtual RFLP pattern derived from the query 16S rDNA fragment of MTWB phytoplasma is most similar to the reference pattern of the 16Sr group I, subgroup B (OY-M, GenBank accession AP006628), with a pattern similarity coefficient of 0.99. The phytoplasma is identified as \'Ca. Phytoplasma asteris\'-related strain belonging to sub-group 16SrI-B. The phylogenetic tree was constructed based on 16S rRNA gene and rp gene sequences by using MEGA version 6.0 (Tamura et al. 2013) with neighbor-joining (NJ) method and bootstrap support was estimated with 1000 replicates. The result indicated that the MTWB phytoplasma formed a subclade in 16SrI-B and rpI-B respectively. In 2013, Macadamia nut showed leaf hardness phyllody and shoot proliferation caused by \'Ca. Phytoplasma asteris\' in Artemisa, Cuba. The concern is that, the macadamia phytoplasma is closely related to the subgroup 16SrI-F, and it is significantly different from the Chinese strains (Pérez-López et al. 2013). In addition, the MTWB phytoplasma was graft-transmitted from infected to healthy plants in nursery conditions (Akhtar et al. 2009; Ikten et al. 2014). And the grafted plants were positive for the phytoplasma in the nested PCR assays. It is noteworthy that the plants were seriously damaged by aphid and it was speculated that the insects of Homoptera caused the spread of the disease by sucking plant sap, thus the aphids that transmits MTWB in China must be determined to control the M. ternifolia witches\'-broom disease. To the best of our knowledge, Macadamia nut is a new host plant of \'Ca. Phytoplasma asteris\' in China. The newly emerged disease is a threat to Macadamia nut.
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    胡桃木(铁核桃)主要分布在中国西南部,并对木材和坚果有价值(Feng等人。2018)。2020年4月,我们调查了位于白沙镇的核桃园,万源市,(四川,中国),在100种植物中的10种树木的叶子上观察到褐斑症状,当树木严重感染时,这种疾病会导致生长潜力降低。在病叶上观察到带有分生孢子的坏死性和亚圆形病变。收集有症状的叶子,并带回实验室进行进一步分析。使用Chomnunti等人开发的单孢子分离技术。(2014),在马铃薯葡萄糖琼脂培养基(PDA)上从感染的叶子上生长了五个分离株。五个分离株具有相似的菌落形态,最初是白色的,近圆形,逐渐变黄,有黑点,发育蓬松的气生菌丝体。在PDA上使用光学显微镜检查形态特征。分生孢子细胞呈亚圆柱形至圆柱形,或者壶腹状,透明玻璃,很少分枝。大分生孢子是月肉,肾形,透明玻璃,1-3-隔膜,主要是1-隔膜,在隔膜处明显收缩,基底细胞是钝圆的,顶端细胞有一个急性末端,基底细胞等于或大于顶端细胞,测量22至40.5×2.5至8.3μm(平均值=32×6.2μm,n=50)。微分生孢子是肉毒杆菌,或亚梭形,透明玻璃,两端都是圆形的,直的或弯曲的,无菌,测量为10至28.5×1.9至3.7μm(平均值=17.2×2.7μm,n=20)。进行多位点方法以精确鉴定代表性分离株SICAUCC20-0012。内部转录间隔区(ITS),鸟嘌呤核苷酸结合蛋白亚基β基因(MS204),分离株SICAUCC20-0012的翻译延伸因子1-α(tef1-α)被扩增并测序,如Sogonov等人所述。(2008)和Walker等人。(2012a)。GenBank登录号对于ITS,MS204和tef1-α分别是MW250303、MW246773和MW246775。系统发育分析显示,蛇形菌具有100%的支持(Fr。)索戈诺夫,形态与Walker等人记录的无性阶段一致。(2012b)。为了测试科赫的假设,如Desai等人所述,在用小别针受伤后,用分生孢子悬浮液(104分生孢子/mL)接种每株植物5-8叶的西吉拉塔(2至3岁)。(2019),相同数量的健康植物受伤,并喷洒无菌蒸馏水作为对照。每天定期用蒸馏水喷洒植物,并将其置于25℃的生长室中,采用12小时的荧光/黑暗方案。15天后,在接种的叶子上出现典型的褐斑症状,但不是控制。如上所述,将真菌O.leptostyla从病变中重新分离出来,但未从未接种的叶子中分离出来。O.在一些核桃树上报道了leptostyla;例如:J.ailantifolia,J.加州,J.Cinerea,J.印度教,J.少校,J.Mandshurica,J.Nigra,和J.Regia(Farr&Rossman2020)。然而,根据我们的知识,这是O.leptostyla在J.sigillata上引起褐色叶斑病的第一份报告。西吉拉塔是中国西南地区的重要经济树种,和杀真菌剂处理应考虑防止这种真菌传播之前,它变得更广泛。杨春林,YuDeng,王飞虎对这项工作做出了同样的贡献。本研究得到四川省重点研发项目(2021YFYZ0032)的资助。
    Juglans sigillata Dode (Iron walnut) is mostly distributed in southwestern China, and valued for wood and nuts (Feng et al. 2018). In April 2020, we surveyed a walnut garden located in Baisha Town, Wanyuan City, (Sichuan, China), where brown spot symptoms were observed on leaves of ten trees among of 100 plants, and this disease can result in a reduced growth potential when trees are severely infected. Necrotic and subcircular lesions with conidiamata were observed on diseased leaves. Symptomatic leaves were collected and taken back to the laboratory forfurther analysis. Using the single spore isolation technique developed by Chomnunti et al. (2014), five isolates were grown from the infected leaves on Potato Dextrose Agar medium (PDA). The five isolates had similar colony morphology, which was initially white, suborbicular, gradually turning yellowish with black spots, developing fluffy aerial mycelium. Morphological characteristics were examined using light microscopy on the PDA. Conidiogenous cells were subcylindrical to cylindrical, or ampulliform, hyaline, rarely branched. Macroconidia were lunate, reniform, hyaline, 1-3-septate, mostly 1-septate, distinctly constricted at the septum, the basal cell was bluntly rounded, the apical cell had an acute end, and the basal cell was equal to or larger than the apical cell, measuring 22 to 40.5 × 2.5 to 8.3 μm (mean = 32 × 6.2 μm, n = 50). Microconidia were botuliform, or subfusiform, hyaline, both ends were rounded, straight or curved, aseptate, and measured 10 to 28.5 × 1.9 to 3.7 μm (mean= 17.2 × 2.7 μm, n = 20). A multilocus approach was conducted for precise identification of a representative isolate SICAUCC 20-0012. The internal transcribed spacer regions (ITS), guanine nucleotide-binding protein subunit beta gene (MS204), and translation elongation factor 1-alpha (tef1-α) of isolate SICAUCC 20-0012 were amplified and sequenced as described by Sogonov et al. (2008) and Walker et al. (2012a). GenBank Accession Nos. for ITS, MS204, and tef1-α are MW250303, MW246773, and MW246775, respectively. Phylogenetic analyses showed 100% support with Ophiognomonia leptostyla (Fr.) Sogonov, and the morphology was consistent with the asexual stage of O. leptostyla documented by Walker et al. (2012b). To test Koch\'s postulates, five healthy plants of J. sigillata (2- to 3-year-old) with 5-8 leaves per plant were inoculated with conidial suspensions (104 conidia/mL) after wounded with a small pin as described by Desai et al. (2019), and the same number of healthy plants were wounded and sprayed with sterile distilled water as controls. Plants were sprayed regularly with distilled water every day and placed in a growth chamber at 25℃ with a 12-h fluorescent light/dark regimen. After 15 days, typical brown spot symptoms developed on inoculated leaves, but not on the controls. The fungus O. leptostyla was reisolated from the lesion as described above but not from non-inoculated leaves. O. leptostyla has been reported on some walnut trees; for example: J. ailantifolia, J. californica, J. cinerea, J. hindsii, J. major, J. mandshurica, J. nigra, and J. regia (Farr & Rossman 2020). However, to our knowledge, this is the first report of O. leptostyla causing brown leaf spot on J. sigillata. J. sigillata is an economically important tree in southwest China, and fungicide treatments should be considered to prevent the spread of this fungus before it becomes more widespread. Chunlin Yang, Yu Deng, and Feihu Wang contributed equally to this work. This research was supported by the Key Research and Development Project of Sichuan Province (2021YFYZ0032).
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