Transfer of transgenes for resistance to rice tungro disease into high-yielding rice cultivars through gene-based marker-assisted selection

SUMMARYRice tungro disease (RTD), caused by the simultaneous infection of rice tungro bacilliform virus (RTBV) and rice tungro spherical virus (RTSV), is one of the major threats to sustainable rice production in South and Southeast Asia. Transgenic resistance against RTBV has been reported previously using an RNA interference (RNAi) construct (ORF IV of RTBV, placed both in sense and anti-sense orientation under CaMV 35S promoter), in the scented rice line Pusa Basmati-1 (PB-1). This construct was transferred to two high-yielding tungro-susceptible indica rice cultivars (IET4094 and IET4786) from the transgenic PB-1 rice line using back cross breeding till the BC2F3 stage. On challenge inoculation, the progenies (BC2F1) showed mild symptoms of tungro, in contrast to severe symptoms displayed by the recurrent parents. Segregation of the transgene indicated near homozygosity of the plants at the BC2F3 stage, implying that the lines can be used as a valuable resistance source for further breeding against RTD.

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Near Immunity to Rice Tungro Spherical Virus Achieved in Rice by a Replicase-Mediated Resistance Strategy

Phytopathology ◽

10.1094/phyto.1999.89.11.1022 ◽

1999 ◽

Vol 89 (11) ◽

pp. 1022-1027 ◽

Cited By ~ 30

Author(s):

H. Huet ◽

S. Mahendra ◽

J. Wang ◽

E. Sivamani ◽

C. A. Ong ◽

...

Keyword(s):

Antisense Rna ◽

Rice Tungro Bacilliform Virus ◽

Japonica Rice ◽

Rice Tungro Spherical Virus ◽

Rice Plants ◽

Antisense Orientation ◽

Rice Tungro Disease ◽

Sense Orientation ◽

Moderate Resistance ◽

High Level

Rice tungro disease is caused by rice tungro bacilliform virus (RTBV), which is responsible for the symptoms, and rice tungro spherical virus (RTSV), which assists transmission of both viruses by leafhoppers. Transgenic japonica rice plants (Oryza sativa) were produced containing the RTSV replicase (Rep) gene in the sense or antisense orientation. Over 70% of the plants contained one to five copies of the Rep gene, with integration occurring at a single locus in most cases. Plants producing antisense sequences exhibited significant but moderate resistance to RTSV (60%); accumulation of antisense RNA was substantial, indicating that the protection was not of the homology-dependent type. Plants expressing the full-length Rep gene, as well as a truncated Rep gene, in the (+)-sense orientation were 100% resistant to RTSV even when challenged with a high level of inoculum. Accumulation of viral RNA was low, leading us to conclude that RTSV Rep-mediated resistance is not protein-mediated but is of the cosuppression type. Resistance was effective against geographically distinct RTSV isolates. In addition, RTSV-resistant transgenic rice plants were unable to assist transmission of RTBV. Such transgenic plants could be used in an epidemiological approach to combat the spread of the tungro disease.

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Characterization of Oryza rufipogon–Derived Resistance to Tungro Disease in Rice

Plant Disease ◽

10.1094/pdis-91-11-1386 ◽

2007 ◽

Vol 91 (11) ◽

pp. 1386-1391 ◽

Cited By ~ 19

Author(s):

Yuji Shibata ◽

Rogelio C. Cabunagan ◽

Pepito Q. Cabauatan ◽

Il-Ryong Choi

Keyword(s):

Interspecific Hybridization ◽

Resistance Mechanisms ◽

Oryza Rufipogon ◽

Rice Tungro Bacilliform Virus ◽

Rice Cultivars ◽

Susceptible Parent ◽

Rice Tungro Spherical Virus ◽

Rice Tungro Disease ◽

Apparent Resistance

Rice tungro disease (RTD) is caused by the interaction between Rice tungro spherical virus (RTSV) and Rice tungro bacilliform virus (RTBV), both of which are transmitted by green leafhoppers (GLH). In order to define the resistance against RTD in rice cv. Matatag 9 which was developed by interspecific hybridization between RTD-susceptible cv. IR64 and Oryza rufipogon, the reactions of Matatag 9 to the viruses and GLH were evaluated in comparison with RTD-susceptible and -resistant rice cultivars. The incidences of infection with RTSV and RTBV in Matatag 9 were significantly lower than those in the susceptible parent cv. IR64; however, no substantial differences in virus accumulation were observed between IR64 and Matatag 9 once infected with the viruses. Symptoms in Matatag 9 infected with RTBV and RTSV were milder than those observed in IR64. A higher level of antixenosis to GLH was observed in Matatag 9 compared with IR64. The levels of antibiosis against GLH in Matatag 9 were comparable with those in another GLH-resistant cultivar, and significantly higher than those in RTD-susceptible cultivars. Collectively, these results suggest that tolerance to tungro viruses and resistance to GLH both contribute to the apparent resistance to RTD in Matatag 9, although possible involvement of other resistance mechanisms cannot be excluded.

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Single Nucleotide Polymorphisms in a Gene for Translation Initiation Factor (eIF4G) of Rice (Oryza sativa) Associated with Resistance to Rice tungro spherical virus

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-23-1-0029 ◽

2010 ◽

Vol 23 (1) ◽

pp. 29-38 ◽

Cited By ~ 52

Author(s):

Jong-Hee Lee ◽

Muhammad Muhsin ◽

Genelou A. Atienza ◽

Do-Yeon Kwak ◽

Suk-Man Kim ◽

...

Keyword(s):

Translation Initiation ◽

Recessive Gene ◽

Translation Initiation Factor ◽

Initiation Factor ◽

Rice Tungro Bacilliform Virus ◽

Nucleotide Polymorphisms ◽

Rice Tungro Spherical Virus ◽

Single Nucleotide ◽

Rice Tungro Disease ◽

Backcross Populations

Rice tungro disease (RTD) is a serious constraint to rice production in South and Southeast Asia. RTD is caused by Rice tungro spherical virus (RTSV) and Rice tungro bacilliform virus. Rice cv. Utri Merah is resistant to RTSV. To identify the gene or genes involved in RTSV resistance, the association of genotypic and phenotypic variations for RTSV resistance was examined in backcross populations derived from Utri Merah and rice germplasm with known RTSV resistance. Genetic analysis revealed that resistance to RTSV in Utri Merah was controlled by a single recessive gene (tsv1) mapped within an approximately 200-kb region between 22.05 and 22.25 Mb of chromosome 7. A gene for putative translation initiation factor 4G (eIF4Gtsv1) was found in the tsv1 region. Comparison of eIF4Gtsv1 gene sequences among susceptible and resistant plants suggested the association of RTSV resistance with one of the single nucleotide polymorphism (SNP) sites found in exon 9 of the gene. Examination of the SNP site in the eIF4Gtsv1 gene among various rice plants resistant and susceptible to RTSV corroborated the association of SNP or deletions in codons for Val1060-1061 of the predicted eIF4Gtsv1 with RTSV resistance in rice.

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Suppression of Two Tungro Viruses in Rice by Separable Traits Originating from Cultivar Utri Merah

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-22-10-1268 ◽

2009 ◽

Vol 22 (10) ◽

pp. 1268-1281 ◽

Cited By ~ 15

Author(s):

Jaymee R. Encabo ◽

Pepito Q. Cabauatan ◽

Rogelio C. Cabunagan ◽

Kouji Satoh ◽

Jong-Hee Lee ◽

...

Keyword(s):

Serological Test ◽

Gene Expression Pattern ◽

Global Gene Expression ◽

Rice Tungro Bacilliform Virus ◽

Near Isogenic Lines ◽

Rice Tungro Spherical Virus ◽

Chain Reaction ◽

Rice Tungro Disease ◽

Polymerase Chain ◽

The Individual

Rice tungro disease (RTD) is caused by Rice tungro spherical virus (RTSV) and Rice tungro bacilliform virus (RTBV) transmitted by green leafhoppers. Rice cv. Utri Merah is highly resistant to RTD. To define the RTD resistance of Utri Merah, near-isogenic lines (NIL, BC5 or BC6) developed from Utri Merah and susceptible cv. Taichung Native 1 (TN1) were evaluated for reactions to RTSV and RTBV. TW16 is an NIL (BC5) resistant to RTD. RTBV was able to infect both TN1 and TW16 but the levels of RTBV were usually significantly lower in TW16 than in TN1. Infection of RTSV was confirmed in TN1 by a serological test but not in TW16. However, the global gene-expression pattern in an RTSV-resistant NIL (BC6), TW16-69, inoculated with RTSV indicated that RTSV can also infect the resistant NIL. Infection of RTSV in TW16 was later confirmed by reverse-transcription polymerase chain reaction but the level of RTSV was considerably lower in TW16 than in TN1. Examination for virus accumulation in another NIL (BC6), TW16-1029, indicated that all plants of TW16-1029 were resistant to RTSV, whereas the resistance to RTBV and symptom severity were segregating among the individual plants of TW16-1029. Collectively, these results suggest that RTD resistance of Utri Merah involves suppression of interacting RTSV and RTBV but the suppression trait for RTSV and for RTBV is inherited separately.

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Engineering Plant Resistance to Tomato Yellow Leaf Curl Thailand Virus Using a Phloem-Specific Promoter Expressing Hairpin RNA

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-06-19-0158-r ◽

2020 ◽

Vol 33 (1) ◽

pp. 87-97

Author(s):

Yuh Tzean ◽

Ho-Hsiung Chang ◽

Tsui-Chin Tu ◽

Bo-Han Hou ◽

Ho-Ming Chen ◽

...

Keyword(s):

Chlorophyll Synthesis ◽

Tomato Yellow Leaf ◽

Cauliflower Mosaic Virus ◽

Rice Tungro Bacilliform Virus ◽

35S Promoter ◽

Yellow Leaf ◽

Hairpin Rna ◽

Leaf Curl Disease ◽

Inoculation Assay ◽

Leaf Curl

Transgenic approaches employing RNA interference (RNAi) strategies have been successfully applied to generate desired traits in plants; however, variations between RNAi transgenic siblings and the ability to quickly apply RNAi resistance to diverse cultivars remain challenging. In this study, we assessed the promoter activity of a cauliflower mosaic virus 35S promoter (35S) and a phloem-specific promoter derived from rice tungro bacilliform virus (RTBV) and their efficacy to drive RNAi against the endogenous glutamate-1-semialdehyde aminotransferase gene (GSA) that acts as a RNAi marker, through chlorophyll synthesis inhibition, and against tomato yellow leaf curl Thailand virus (TYLCTHV), a begomovirus (family Geminiviridae) reported to be the prevalent cause of tomato yellow leaf curl disease (TYLCD) in Taiwan. Transgenic Nicotiana benthamiana expressing hairpin RNA of GSA driven by either the 35S or RTBV promoter revealed that RTBV::hpGSA induced stronger silencing along the vein and more uniformed silencing phenotype among its siblings than 35S::hpGSA. Analysis of transgenic N. benthamiana, 35S::hpTYLCTHV, and RTBV::hpTYLCTHV revealed that, although 35S::hpTYLCTHV generated a higher abundance of small RNA than RTBV::hpTYLCTHV, RTBV::hpTYLCTHV transgenic plants conferred better TYLCTHV resistance than 35S::hpTYLCTHV. Grafting of wild-type (WT) scions to TYLCTHV RNAi rootstocks allowed transferable TYLCTHV resistance to the scion. A TYLCTHV-inoculation assay showed that noninfected WT scions were only observed when grafted to RTBV::hpTYLCTHV rootstocks but not 35S::hpTYLCTHV nor WT rootstocks. Together, our findings demonstrate an approach that may be widely applied to efficiently confer TYLCD resistance.

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Whole Genome Sequencing of Elite Rice Cultivars as a Comprehensive Information Resource for Marker Assisted Selection

PLoS ONE ◽

10.1371/journal.pone.0124617 ◽

2015 ◽

Vol 10 (4) ◽

pp. e0124617 ◽

Cited By ~ 42

Author(s):

Jorge Duitama ◽

Alexander Silva ◽

Yamid Sanabria ◽

Daniel Felipe Cruz ◽

Constanza Quintero ◽

...

Keyword(s):

Whole Genome Sequencing ◽

Genome Sequencing ◽

Marker Assisted Selection ◽

Information Resource ◽

Whole Genome ◽

Rice Cultivars ◽

Comprehensive Information

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Transformation of Carrot (Daucus carota L.) with Genes Involved in Carbohydrate Metabolism and Partitioning via Hypocotyl Tissue Cultures

HortScience ◽

10.21273/hortsci.32.3.453e ◽

1997 ◽

Vol 32 (3) ◽

pp. 453E-453

Author(s):

Mingbo Qin ◽

Chiwon W. Lee ◽

Alex Y. Borovkov ◽

Murray E. Duysen

Keyword(s):

Daucus Carota ◽

Sugar Metabolism ◽

Sucrose Phosphate Synthase ◽

Kanamycin Resistance ◽

35S Promoter ◽

Daucus Carota L ◽

Sense Orientation ◽

Asexual Embryogenesis ◽

Hypocotyl Tissue ◽

Root Tissues

A study was initiated to characterize key enzymes that influence sweetness in carrot (Daucus carota L.) roots. Sucrose synthase (SS), sucrose phosphate synthase (SPS), and UDP-glucose pyrophosphorylase (UDPL) genes were isolated from potato (Solanum tuberosum L.) and cloned in an anti-sense orientation into Agrobacterium tumefaciens Bin19, which has a CaMV 35S promoter. Seedling hypocotyl sections of selected carrot lines were pre-incubated on B5 medium for 2 days, co-cultivated with A. tumefaciens Bin 19 for additional 3 days, and then transferred to a modified B5 medium containing 50 g/mL kanamycin and 400 g/mL carbenicillin. In 4 weeks, 18.6%, 33.3%, and 26.7% of the cultures from a breeding line (W204-C) were found to be transformed, respectively, with SS, SPS, and UDPL as determined by kanamycin resistance. In contrast, no kanamycin-resistant calli were obtained from a commercial cultivar (Navajo) in these transformation studies. The transformed calli proliferated in the medium containing 50 g/mL kanamycin and 400 g/mL carbenicillin, whereas non-transformed calli died in the same medium. These transformed calli are currently being used to regenerate plants via asexual embryogenesis using a suspension culture. The influence of these additional genes on sugar metabolism and accumulation in root tissues of transformed carrots will be characterized in the future.

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The sense and antisense coat protein gene of alfalfa mosaic virus strain N20 confers protection in transgenic tobacco plants

Australian Journal of Agricultural Research ◽

10.1071/a96098 ◽

1997 ◽

Vol 48 (4) ◽

pp. 503 ◽

Cited By ~ 3

Author(s):

K. W. Jayasena ◽

B. J. Ingham ◽

M. R. Hajimorad ◽

J. W. Randles

Keyword(s):

Coat Protein ◽

Mosaic Virus ◽

Coat Protein Gene ◽

Protein Gene ◽

Alfalfa Mosaic Virus ◽

35S Promoter ◽

Pasture Legumes ◽

Nicotiana Tabacum L ◽

Sense Orientation ◽

A Minor

The coat protein gene of a South Australian strain of alfalfa mosaic virus (AMV-N20 [NcS]) has been cloned, sequenced, and transferred into Nicotiana tabacum L. cv. Xanthi via Agrobacterium tumefaciens under the control of the CaMV 35S promoter. A number of lines (T0 generation) were selected with the coat protein gene either in sense orientation (CP+) or in antisense orientation (CP–). The T0 plants were tested for their gene expression and susceptibility to the homologous AMV strain. A significant delay in the onset of symptoms and a reduction in virus accumulation was observed in CP+ plants mechanically inoculated with AMV. CP– plants were also significantly protected but less so than the CP+ plants. Plants transformed with the expression vector only (CP0) showed a minor resistance to local infection on inoculated leaves compared with untransformed plants. The strategy of coat protein mediated protection (CPMP) using the CP gene in either messenger sense or antisense would therefore be appropriate for testing on economically important pasture legumes.

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Assessment of resistance to rice tungro disease in popular rice varieties in India by introgression of a transgene against Rice tungro bacilliform virus

Archives of Virology ◽

10.1007/s00705-019-04159-3 ◽

2019 ◽

Vol 164 (4) ◽

pp. 1005-1013 ◽

Cited By ~ 1

Author(s):

G. Kumar ◽

M. Jyothsna ◽

P. Valarmathi ◽

S. Roy ◽

A. Banerjee ◽

...

Keyword(s):

Rice Tungro Bacilliform Virus ◽

Rice Varieties ◽

Rice Tungro Disease

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Complete Genome Sequence of Rice Tungro Bacilliform Virus Infecting Asian Rice (Oryza sativa) in Malaysia

Microbiology Resource Announcements ◽

10.1128/mra.00262-19 ◽

2019 ◽

Vol 8 (20) ◽

Author(s):

Maathavi Kannan ◽

Maisarah Mohamad Saad ◽

Noraini Talip ◽

Syarul Nataqain Baharum ◽

Hamidun Bunawan

Keyword(s):

Oryza Sativa ◽

Genome Sequence ◽

Complete Genome Sequence ◽

Complete Genome ◽

Rice Tungro Bacilliform Virus ◽

Asian Rice ◽

Rice Tungro Disease

Rice tungro disease was discovered in Malaysia in the 1930s. The first and only genome of Rice tungro bacilliform virus (RTBV) isolated from rice in Malaysia was sequenced in 1999.

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