Scholarlink

<p><i>Xanthomonas oryzae</i> pv. <i>oryzae</i> (<i>Xoo</i>) is a destructive pathogen that causes bacterial leaf blight (BLB) disease in rice. Along with <i>Xoo</i>, the BLB-infected leaves are inhabited by many bacterial species, which all produce morphologically similar colonies on culture media, making the selection of <i>Xoo</i> difficult. In the present study, we have tested a simple and modified streaking technique, i.e., the needle-stabbing-streaking method, for rapid and accurate isolation of <i>Xoo</i>. We have used seven different culture media, namely growth factor agar (GFA), nutrient broth yeast extract agar (NBYEA), peptone sucrose agar (PSA), yeast dextrose chalk agar (YDCA), modified <i>Xanthomonas oryzae</i> specific media (mXOS), modified Wakimoto’s medium (MWM), and nutrient agar (NA). We have standardized and adopted a colony polymerase chain reaction (colony PCR) assay for taxonomic confirmation using previously reported <i>Xoo</i>-specific primers. We also employed two other conventional techniques, ooze streaking and direct leaf streaking, for comparison. Among the different methods tested, the needle-stabbing-streaking method provided the maximum <i>Xoo</i> colony count on all the media; however, maximum colonies were recorded on the PSA (<i>n</i> = 36), NBYEA (<i>n</i> = 35), and NA (<i>n</i> = 30) with less or zero contamination on all media within 4 days after inoculation. We also tested the effectiveness of each method for isolating <i>Xoo</i> from the stored samples, which indicated that the needle-stabbing-streaking method could yield positive <i>Xoo</i> colonies up to 15 days after sampling, suggesting its potential wider applicability. Further, this method was validated using on-field-collected BLB-infected leaf samples of three commercial rice cultivars. Therefore, the needle-stabbing-streaking is simple, rapid, efficient, and cultivar-media-independent for the accurate isolation of <i>Xoo</i>.</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

Needle-stabbing-streaking: a simple and accurate technique for rapid isolation of Xanthomonas oryzae pv. oryzae from rice leaves

  • 期刊论文

  • 发表日期：

    2026年3月19日

查看全文
  • Manne Tulasi,
  • Devanna Pramesh,
  • Mukanara Nagarajappa Harish,
  • Muthakapalli Krishnareddy Prasannakumar,
  • Channappa Manjunatha

摘要

Xanthomonas oryzae pv. oryzae (Xoo) is a destructive pathogen that causes bacterial leaf blight (BLB) disease in rice. Along with Xoo, the BLB-infected leaves are inhabited by many bacterial species, which all produce morphologically similar colonies on culture media, making the selection of Xoo difficult. In the present study, we have tested a simple and modified streaking technique, i.e., the needle-stabbing-streaking method, for rapid and accurate isolation of Xoo. We have used seven different culture media, namely growth factor agar (GFA), nutrient broth yeast extract agar (NBYEA), peptone sucrose agar (PSA), yeast dextrose chalk agar (YDCA), modified Xanthomonas oryzae specific media (mXOS), modified Wakimoto’s medium (MWM), and nutrient agar (NA). We have standardized and adopted a colony polymerase chain reaction (colony PCR) assay for taxonomic confirmation using previously reported Xoo-specific primers. We also employed two other conventional techniques, ooze streaking and direct leaf streaking, for comparison. Among the different methods tested, the needle-stabbing-streaking method provided the maximum Xoo colony count on all the media; however, maximum colonies were recorded on the PSA (n = 36), NBYEA (n = 35), and NA (n = 30) with less or zero contamination on all media within 4 days after inoculation. We also tested the effectiveness of each method for isolating Xoo from the stored samples, which indicated that the needle-stabbing-streaking method could yield positive Xoo colonies up to 15 days after sampling, suggesting its potential wider applicability. Further, this method was validated using on-field-collected BLB-infected leaf samples of three commercial rice cultivars. Therefore, the needle-stabbing-streaking is simple, rapid, efficient, and cultivar-media-independent for the accurate isolation of Xoo.