Soto-Heredia José, Ramos-Tito Salome, Alves Angelica Rodrigues, Ferreira Luciellen da Costa, Calderon Luz Leonor Mattos, Rossato Maurício
Universidade de Brasilia, Plant Pathology, Brasilia, DF, Brazil.
Universidad Nacional Agraria La Molina, Plant Pathology, La Molina, Peru;
Plant Dis. 2024 Oct 9. doi: 10.1094/PDIS-09-24-1874-PDN.
Around 90% of Peru's ginger (Zingiber officinale) production is concentrated in the Junín region, due to the optimal agroecological conditions for its cultivation. In March 2024, fields with ginger plants (cultivar Criollo) in Junín region, provinces Chanchamayo and Satipo, specifically in the cities of Pichanaqui and Satipo respectively, exhibited approximately 40% of plants with severe symptoms of a disease characterized initially by plant yellowing and rapid progressing to necrosis. Affected rhizomes showed dark vascular bundles with milky white exudates upon cutting, while stems displayed vascular necrosis hindering water and nutrient transport, often resulting in plant death. Fifteen plants were sampled and diseased vascular tissues from rhizomes and stems were cultured on nutrient agar (NA) and incubated at 28°C. After 72 h, all isolations resulted in colonies with typical characteristics of Ralstonia solanacearum species complex (RSSC) were produced, with appearing fluid, irregularly round, and creamy white. Three isolates were selected for the identification steps (UNALM-RP01 to 03) were identified by PCR using primers 759/760 (Opina et al. 1997) confirming as RSSC with a 282 bp amplification product. Additionally, isolates were assigned to biovar 3 based on their ability to metabolize three acid-producing disaccharides (maltose, lactose, cellobiose) and three hexose alcohols (mannitol, sorbitol, dulcitol) (Hayward, 1964). Phylotype I was identified by multiplex PCR (primers Nmult) with a 114 bp amplification product (Fegan and Prior 2005). For the identification of the sequevars of the three isolates, DNA was extracted and PCR with primers ENDO-F/R (Ji et al. 2007) were performed to amplify and sequence the partial gene sequence of egl gene with 681 bp in length. The phylogeny by Neighbor joining with 10,000 bootstraps clustered the UNALM isolates along other sequevar 30 of R. pseudosolanacearum. The sequences were deposited in Genbank under accessions PQ213016, PQ213017 and PQ213018. For pathogenicity tests, bacterial colonies of isolate UNALM-RP01 were scraped from the culture media with a sterile needle and introduced into the stems of three 2-month-old ginger plants (cultivar Gigante). The plants subsequently exhibited yellowing seven days post-inoculation. Additionally, the rhizomes showed internal discoloration and bacterial exudation. Three plants were used as a control, which were pierced with a sterilized needle and showed no symptoms. All tested plants were kept in a greenhouse with controlled temperature (20-40 °C) The pathogen was successfully re-isolated from infected plants on NA medium, presenting typical colonies of RSSC and identified via PCR with primers 759/760, fulfilling Koch's postulates. This represents the first case in Peru of ginger plants infected with a Ralstonia species, specifically R. solanacearum phylotype I, corresponding to R. pseudosolanacearum. This species of RSSC and sequevar is known for causing disease in ginger. Its presence in Peru, however, may be the result of the pathogen's introduction, as its geographical origin is associated with Asia (Fegan and Prior 2005). To our knowledge, this is the first report of R. pseudosolanacearum causing ginger wilt disease in Peru. In 2024, an estimated average yield loss of 30% has been attributed to wilt disease in the Junín region, posing a significant threat to cultivation. Urgent and effective disease management strategies are essential to control and mitigate further losses.
由于生姜(姜科姜属)种植的最佳农业生态条件,秘鲁约90%的生姜产量集中在胡宁地区。2024年3月,胡宁地区、昌查马约省和萨蒂波省种植生姜植株(品种克里奥洛)的田地,具体分别在皮查纳基市和萨蒂波市,约40%的植株表现出一种疾病的严重症状,最初表现为植株发黄,并迅速发展为坏死。受影响的根茎切割后显示出深色维管束和乳白色渗出物,而茎部则显示维管束坏死,阻碍水分和养分运输,常导致植株死亡。采集了15株植株,将根茎和茎部患病的维管组织在营养琼脂(NA)上培养,并在28°C下孵育。72小时后,所有分离培养均产生了具有茄科劳尔氏菌复合种(RSSC)典型特征的菌落,菌落呈流体状、不规则圆形且乳白色。选择了三个分离株进行鉴定步骤(UNALM - RP01至03),使用引物759/760通过PCR进行鉴定(奥皮纳等人,1997年),确认其为RSSC,扩增产物为282 bp。此外,根据分离株代谢三种产酸二糖(麦芽糖、乳糖、纤维二糖)和三种己糖醇(甘露醇、山梨醇、卫矛醇)的能力,将其归为生物型3(海沃德,1964年)。通过多重PCR(引物Nmult)鉴定出系统型I,扩增产物为114 bp(费根和普赖尔,2005年)。为了鉴定这三个分离株的序列变种,提取DNA并使用引物ENDO - F/R进行PCR(季等人,2007年),以扩增并测序长度为681 bp的egl基因部分序列。通过邻接法构建的系统发育树,经10000次重复抽样,将UNALM分离株与其他假茄劳尔氏菌序列变种30聚类在一起。这些序列已保存在Genbank中,登录号分别为PQ213016、PQ213017和PQ213018。为了进行致病性测试,用无菌针从培养基上刮下分离株UNALM - RP01的细菌菌落,并接种到三株2个月大的生姜植株(品种巨人)的茎中。接种后七天,植株出现发黄。此外,根茎显示内部变色和细菌渗出。使用三株植株作为对照,用消毒针穿刺,未出现症状。所有测试植株均保存在温度可控的温室中(20 - 40°C)。病原体成功地从感染植株上在NA培养基上重新分离出来,呈现出RSSC的典型菌落,并通过引物759/760进行PCR鉴定,符合柯赫氏法则。这是秘鲁首次出现生姜植株感染劳尔氏菌属物种的案例,具体为I型茄科劳尔氏菌,对应假茄劳尔氏菌。这种RSSC物种和序列变种以导致生姜病害而闻名。然而,它在秘鲁的出现可能是病原体引入的结果,因为其地理起源与亚洲有关(费根和普赖尔,2005年)。据我们所知,这是秘鲁首次报道假茄劳尔氏菌引起生姜枯萎病。2024年,据估计胡宁地区因枯萎病造成的平均产量损失达30%,对种植构成了重大威胁。迫切需要有效的病害管理策略来控制和减轻进一步的损失。