Tal2c Activates the Expression of OsF3H04g to Promote Infection as a Redundant TALE of Tal2b in Xanthomonas oryzae pv. oryzicola

Xanthomonas oryzae delivers transcription activator-like effectors (TALEs) into plant cells to facilitate infection. Following economic principles, the redundant TALEs are rarely identified in Xanthomonas. Previously, we identified the Tal2b, which activates the expression of the rice 2-oxoglutarate-dependent dioxygenase gene OsF3H03g to promote infection in the highly virulent strain of X. oryzae pv. oryzicola HGA4. Here, we reveal that another clustered TALE, Tal2c, also functioned as a virulence factor to target rice OsF3H04g, a homologue of OsF3H03g. Transferring Tal2c into RS105 induced expression of OsF3H04g to coincide with increased susceptibility in rice. Overexpressing OsF3H04g caused higher susceptibility and less salicylic acid (SA) production compared to wild-type plants. Moreover, CRISPR–Cas9 system-mediated editing of the effector-binding element in the promoters of OsF3H03g or OsF3H04g was found to specifically enhance resistance to Tal2b- or Tal2c-transferring strains, but had no effect on resistance to either RS105 or HGA4. Furthermore, transcriptome analysis revealed that several reported SA-related and defense-related genes commonly altered expression in OsF3H04g overexpression line compared with those identified in OsF3H03g overexpression line. Overall, our results reveal a functional redundancy mechanism of pathogenic virulence in Xoc in which tandem Tal2b and Tal2c specifically target homologues of host genes to interfere with rice immunity by reducing SA.

KETAHANAN BEBERAPA GENOTIPE PADI HARAPAN TERHADAP PENYAKIT HAWAR DAUN BAKTERI (Xanthomonas oryzae pv. oryzae) STRAIN III, IV DAN VIII.

Penyakit hawar daun bakteri (HDB) yang disebabkan oleh bakteri Xanthomonas oryzae pv. oryzae (Xoo) merupakan penyakit penting yang sering menyerang tanaman padi. Pengujian ketahanan genotipe tanaman terhadap penyakit HBD merupakan sebagai salah satu syarat pelepasan varietas tanaman. Tujuan dari penelitian ini adalah untuk memperoleh informasi potensi genotipe-genotipe padi yang tahan terhadap penyakit HDB yang disebabkan oleh Xanthomonas oryzae pv. oryzae (Xoo) strain III, IV dan VIII di Rumah Kaca. Eksperimen ditata dalam rancangan Split Plot, strain sebagai petak utama dan genotipe sebagai anak petak. Ada sepuluh genotipe padi dan tujuh varietas cek dievaluasi dengan menggunakan analisis varians dan uji lanjut Least Significant Increase (LSI). Hasil penelitian berdasarkan uji lanjut LSI menunjukkan seluruh karakter pengamatan, baik pengamatan periode inkubasi, keparahan penyakit dan jumlah stomata menunjukkan bahwa seluruh genotipe padi (SP101-3-1-5-2, SP101-3-1-5-8, SP101-3-1-19-26, SP101-3-1-38-4, SP87-1-1-7-10, SP101-3-1-19-27, SP101-3-1-38-25, SP87-1-1-7-7, PP48-5-24 dan PP48-5-1) tidak menampilkan perbedaan nyata dengan tetua (Sintanur, Pandanwangi, dan PTB 33) dan varietas cek lainnya (Ciherang, Inpari 32, IRRB7, dan TN1). Semua genotipe uji menunjukkan reaksi rentan pada strain III, dan sangat rentan pada strain IV dan VIII. Pada strain III, baik genotipe uji dan tetua varietas menunjukkan skor lima (rentan), sedangkan strain IV dan strain VIII menunjukkan antara skor enam dan tujuh (sangat rentan). Ditemukan pula beberapa genotipe dengan skor 1 dan 2 (tahan), hal tersebut dapat membuka kemungkinan ditemukannya genotipe yang tahan. Riset ini menunjukkan bahwa komposisi genotipe berpengaruh terhadap reaksi ketahanan terhadap hawar daun bakteri dan terdapat variasi virulensi HBD.

A Pathogen-Inducible Rice NAC Transcription Factor ONAC096 Contributes to Immunity Against Magnaprothe oryzae and Xanthomonas oryzae pv. oryzae by Direct Binding to the Promoters of OsRap2.6, OsWRKY62, and OsPAL1

The rice NAC transcriptional factor family harbors 151 members, and some of them play important roles in rice immunity. Here, we report the function and molecular mechanism of a pathogen-inducible NAC transcription factor, ONAC096, in rice immunity against Magnaprothe oryzae and Xanthomonas oryzae pv. oryzae. Expression of ONAC096 was induced by M. oryzae and by abscisic acid and methyl jasmonate. ONAC096 had the DNA binding ability to NAC recognition sequence and was found to be a nucleus-localized transcriptional activator whose activity depended on its C-terminal. CRISPR/Cas9-mediated knockout of ONAC096 attenuated rice immunity against M. oryzae and X. oryzae pv. oryzae as well as suppressed chitin- and flg22-induced reactive oxygen species burst and expression of PTI marker genes OsWRKY45 and OsPAL4; by contrast, overexpression of ONAC096 enhanced rice immunity against these two pathogens and strengthened chitin- or flg22-induced PTI. RNA-seq transcriptomic profiling and qRT-PCR analysis identified a small set of defense and signaling genes that are putatively regulated by ONAC096, and further biochemical analysis validated that ONAC096 could directly bind to the promoters of OsRap2.6, OsWRKY62, and OsPAL1, three known defense and signaling genes that regulate rice immunity. ONAC096 interacts with ONAC066, which is a positive regulator of rice immunity. These results demonstrate that ONAC096 positively contributes to rice immunity against M. oryzae and X. oryzae pv. oryzae through direct binding to the promoters of downstream target genes including OsRap2.6, OsWRKY62, and OsPAL1.

Antagonistic activity of rice rhizosphere isolates against Xanthomonas oryzae pv. oryzae bacterial blight pathogen

Attempts were made to isolate beneficial bio agents from rice rhizosphere which resulted in isolation of 46 Bacillus spp and 15 fluorescent Pseudomonas spp which were further investigated for their potential aginst BB of rice diseaseAmong twenty six isolates of Bacillus, two isolates were most antagonistic and showed highest inhibition percentage (57.09) The potential isolates of Pseudomonas (P-4, P-5, P-6, P-7 and P-8), Bacillus (ARI 1-2, ARI 2-4, ARI 1-3, D1-1-2 and D2-1-1) were tested in vivo under glass house conditions for antagonism against Xoo which revealed that P-7 was effective in reduction of lesion length (18.5cm) when compared to control (20.37cm). The potential isolates of Pseudomonas (P-4, P-5, P-6, P-7 and P-8), Bacillus (ARI 1-2, ARI 2-4, ARI 1-3, D1-1-2 and D2-1-1) were tested in vivo under glass house conditions for antagonism against Xoo which revealed that P-7 was effective in reduction of lesion length (18.5cm) when compared to control (20.37cm). The present study indicated that PGPR isolates P-4, P-6, P-7 and P-8 can be used as biofertilizers, which will add up for enhanced growth of rice.

Discovery of Novel Dihydrolipoamide S-Succinyltransferase Inhibitors Based on Fragment Virtual Screening

A series of new oxadiazole sulfone derivatives containing an amide moiety was synthesized based on fragment virtual screening to screen high-efficiency antibacterial agents for rice bacterial diseases. All target compounds showed greater bactericidal activity than commercial bactericides. 3-(4-fluorophenyl)-N-((5-(methylsulfonyl)-1,3,4-oxadiazol-2-yl)methyl)acrylamide (10) showed excellent antibacterial activity against Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola, with EC50 values of 0.36 and 0.53 mg/L, respectively, which were superior to thiodiazole copper (113.38 and 131.54 mg/L) and bismerthiazol (83.07 and 105.90 mg/L). The protective activity of compound 10 against rice bacterial leaf blight and rice bacterial leaf streak was 43.2% and 53.6%, respectively, which was superior to that of JHXJZ (34.1% and 26.4%) and thiodiazole copper (33.0% and 30.2%). The curative activity of compound 10 against rice bacterial leaf blight and rice bacterial leaf streak was 44.5% and 51.7%, respectively, which was superior to that of JHXJZ (32.6% and 24.4%) and thiodiazole copper (27.1% and 28.6%). Moreover, compound 10 might inhibit the growth of Xanthomonas oryzae pv. oryzae and Xanthomonas oryzae pv. oryzicola by affecting the extracellular polysaccharides, destroying cell membranes, and inhibiting the enzyme activity of dihydrolipoamide S-succinyltransferase.