Marker‐assisted forward breeding to develop a drought‐, bacterial‐leaf‐blight‐, and blast‐resistant rice cultivar

TBR225 is one of the most popular commercial rice varieties in Northern Vietnam. However, this variety is highly susceptible to bacterial leaf blight (BLB), a disease caused by Xanthomonas oryzae pv. oryzae (Xoo) which can lead to important yield losses. OsSWEET14 belongs to the SWEET gene family that encodes sugar transporters. Together with other Clade III members, it behaves as a susceptibility (S) gene whose induction by Asian Xoo Transcription-Activator-Like Effectors (TALEs) is absolutely necessary for disease. In this study, we sought to introduce BLB resistance in the TBR225 elite variety. First, two Vietnamese Xoo strains were shown to up-regulate OsSWEET14 upon TBR225 infection. To investigate if this induction is connected with disease susceptibility, nine TBR225 mutant lines with mutations in the AvrXa7, PthXo3 or TalF TALEs DNA target sequences of the OsSWEET14 promoter were obtained using the CRISPR/Cas9 editing system. Genotyping analysis of T0 and T1 individuals showed that mutations were stably inherited. None of the examined agronomic traits of three transgene-free T2 edited lines were significantly different from those of wild-type TBR225. Importantly, one of these T2 lines, harboring the largest homozygous 6-bp deletion, displayed decreased OsSWEET14 expression as well as a significantly reduced susceptibility to a Vietnamese Xoo strains and complete resistance to another one. Our findings indicate that CRISPR/Cas9 editing conferred an improved BLB resistance to a Vietnamese commercial elite rice variety.

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Close linkage of a blast resistance gene, Pias(t), with a bacterial leaf blight resistance gene, Xa1-as(t), in a rice cultivar ‘Asominori’

Breeding Science ◽

10.1270/jsbbs.62.334 ◽

2012 ◽

Vol 62 (4) ◽

pp. 334-339 ◽

Cited By ~ 6

Author(s):

Takashi Endo ◽

Masayuki Yamaguchi ◽

Ryota Kaji ◽

Koji Nakagomi ◽

Tomomori Kataoka ◽

...

Keyword(s):

Resistance Gene ◽

Blast Resistance ◽

Rice Cultivar ◽

Leaf Blight ◽

Close Linkage ◽

Bacterial Leaf Blight ◽

Blight Resistance ◽

Blast Resistance Gene

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The Role of Nanochitosan on the Expression of Rice Resistance Genes against Bacterial Leaf Blight

Jurnal Perlindungan Tanaman Indonesia ◽

10.22146/jpti.44418 ◽

2020 ◽

Vol 24 (2) ◽

pp. 115

Author(s):

Siswanti Siswanti ◽

Tri Joko ◽

Siti Subandiyah

Keyword(s):

Resistance Genes ◽

Rapid Evolution ◽

Rice Cultivar ◽

Leaf Blight ◽

Bacterial Leaf Blight ◽

Distilled Water ◽

Disease Symptoms ◽

Resistant Varieties ◽

Significant Difference

Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) has been reported to cause ± 20–50% of rice yield loss around the world. Resistant varieties are used to control this disease, however due to rapid evolution of this pathogen, the resistances was broken down in a few years. This study is aimed to determine the role of nanochitosan in the expression of rice Xa21 and Xa1 resistant genes against Xoo. The BLB susceptible rice cultivar IR64, the Xoo isolate MAG2 and a 0.065% concentration of nanochitosan were used in this experiment. Application of nanochitosan was carried out within 1-week intervals starting at rice aged 2–10 weeks after transplanting. The expression of Xa21 and Xa1 genes against Xoo were analyzed using conventional PCR and qPCR methods at 0 and 4 days after Xoo inoculation followed by 4x scoring of disease symptoms in 1-week interval. The treatments used in this study included the mock one/inoculated with sterile distilled water, K (+)/ plants inoculated with Xoo, CNP (-)/ with nanochitosan and sterile distilled water inoculation, and CNP (+)/ with nanochitosan and Xoo inoculation. The results showed that the 0.0065% concentration nanochitosan application was able to increase the expression of Xa21 and Xa1 genes on CNP (-). Disease intensity and AUDPC values did not show any significant difference between K (+) and CNP (+). This study concluded that nanochitosan at 0.065% was able to increase the expression of rice Xa21 and Xa1 resistance genes. However, the gene expression was not able to significantly suppress the infection development of Xoo.

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Genetic Analysis of Resistance Against Bacterial Leaf Blight and Leaf Blast Disease in the Japanese Rice Cultivar Asominori

Advances in Genetics, Genomics and Control of Rice Blast Disease ◽

10.1007/978-1-4020-9500-9_30 ◽

2009 ◽

pp. 305-313 ◽

Cited By ~ 1

Author(s):

Takashi Endo ◽

Toshiki Nakamura ◽

Junichi Yonemaru ◽

Goro Ishikawa ◽

Masayuki Yamaguchi ◽

...

Keyword(s):

Genetic Analysis ◽

Rice Cultivar ◽

Leaf Blight ◽

Blast Disease ◽

Bacterial Leaf Blight ◽

Leaf Blast

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Transcriptional profiling of indica rice cultivar IET8585 (Ajaya) infected with bacterial leaf blight pathogen Xanthomonas oryzae pv oryzae

Plant Physiology and Biochemistry ◽

10.1016/j.plaphy.2007.07.013 ◽

2007 ◽

Vol 45 (10-11) ◽

pp. 834-850 ◽

Cited By ~ 29

Author(s):

Kameswara Rao Kottapalli ◽

Randeep Rakwal ◽

Kouji Satoh ◽

Junko Shibato ◽

Pratibha Kottapalli ◽

...

Keyword(s):

Indica Rice ◽

Transcriptional Profiling ◽

Rice Cultivar ◽

Leaf Blight ◽

Xanthomonas Oryzae ◽

Bacterial Leaf Blight ◽

Indica Rice Cultivar

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Marker-Assisted Selection of Xa21 Conferring Resistance to Bacterial Leaf Blight in indica Rice Cultivar LT2

Rice Science ◽

10.1016/j.rsci.2017.08.004 ◽

2018 ◽

Vol 25 (1) ◽

pp. 52-56 ◽

Cited By ~ 7

Author(s):

Hue Thi Nguyen ◽

Quang Hong Vu ◽

Tan Van Mai ◽

Thu Thi Nguyen ◽

Lam Duc Vu ◽

...

Keyword(s):

Marker Assisted Selection ◽

Indica Rice ◽

Rice Cultivar ◽

Leaf Blight ◽

Bacterial Leaf Blight ◽

Selection Of ◽

Indica Rice Cultivar

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Design and transfer of OsSWEET14-editing T-DNA construct to bac thom 7 rice cultivar

ACADEMIA JOURNAL OF BIOLOGY ◽

10.15625/2615-9023/14923 ◽

2021 ◽

Vol 43 (1) ◽

Author(s):

Vu Hoai Sam ◽

Pham Thi Van ◽

Nguyen Thanh Ha ◽

Nguyen Thi Thu Ha ◽

Phung Thi Thu Huong ◽

...

Keyword(s):

Rice Variety ◽

Sugar Transport ◽

Rice Cultivar ◽

Leaf Blight ◽

Xanthomonas Oryzae ◽

Bacterial Leaf Blight ◽

Induced Mutations ◽

Transcription Activator ◽

Rna Complex

Bac thom 7 rice (BT7 rice) is one of the major elite rice cultivars in Vietnam with superior productivity and quality but very susceptible to bacterial leaf blight caused by Xanthomonas oryzae pv. oryzae. The gene OsSWEET14, belonging to the OsSWEET family which encodes sugar transport proteins, is considered to be a susceptible gene involved in the virulence of Xoo. At least three cis-elements (EBE - Effector-binding element), including Tal5, PthXo3 and AvrXa7 on BT7 OsSWEET14 promoter, bind specifically to well-known transcription activator–like effectors (TALEs) of many Asian Xoo bacterium strains. In this study, a T-DNA construct which expressed the protein-RNA complex CRISPR/Cas for editing three EBEs position on the OsSWEET14 promoter was designed. The recombinant binary vector was tested by PCR, restriction enzyme and finally sequencing and then successfully transferred to Bac thom 7 rice through Agrobacterium tumefaciens. 28 of 30 hygromycin-resistant regenerated rice lines that grew and developed normally under nethouse conditions were selected by PCR with specific primers. Among these, twelve transgenic rice lines were identified carrying one copy of the T-DNA construct. The presence of CRISPR/Cas9-induced mutations of the targeted promoter in the transgenic plants were confirmed by T7EI assay. These results provide the basis to determine the role of OsSWEET14 in the susceptibility of Bac thom 7 rice to Xanthomonas oryzae pv. oryzae -caused disease, towards the further goal of generating an improved Bac thom 7 rice variety with broad-spectrum bacterial leaf blight resistance using CRISPR/Cas9 technology.

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