I. Oña

Over-expression of the cloned rice thaumatin-like protein (PR-5) gene in transgenic rice plants enhances environmental friendly resistance to Rhizoctonia solani causing sheath blight disease

Abstract A 1.1-kb DNA fragment containing the coding region of a thaumatin-like protein (TLP-D34), a member of the PR-5 group, was cloned into the rice transformation vector pGL2, under the control of the CaMV 35S promoter. The Indica rice cultivars, ‘Chinsurah Boro II’, ‘IR72’, and ‘IR51500’ were transformed with the tlp gene construct by PEG-mediated direct gene transfer to protoplasts and by biolistic transformation using immature embryos. The presence of the chimeric gene in T0, T1, and T2 transgenic plants was detected by Southern blot analysis. The presence of the expected 23-kDa TLP in transgenic plants was confirmed by Western blot analysis and by staining with Coomassie Brilliant Blue. Bioassays of transgenic plants challenged with the sheath blight pathogen, Rhizoctonia solani, indicated that over-expression of TLP resulted in enhanced resistance compared to control plants.

Enhanced resistance to sheath blight by constitutive expression of infection-related rice chitinase in transgenic elite indica rice cultivars.

Genetic transformation has been attempted for management of rice sheath blight disease, caused by Rhizoctonia solani. We introduced a PR-3 rice chitinase gene (RC7), isolated from R. solani-infected rice plants, into indica rice cultivars IR72, IR64, IR68899B, MH63, and Chinsurah Boro II by the biolistic and PEG-mediated transformation system. Inheritance was studied up to the T(2) generation by Southern blot analysis. Western blot analysis of transgenic plants with polyclonal antibody revealed the presence of chitinase protein with a molecular weight of 35 kDa that reacts with chitinase antibody. The transformants synthesized different levels of chitinase proteins constitutively and progeny from the plants containing the chitinase gene showed different levels of enhanced resistance when challenged with the sheath blight pathogen R. solani.

Transgenic rice variety 'IR72' with Xa21 is resistant to bacterial blight

Abstract An elite indica rice variety, ‘IR72’, was transformed with a cloned gene, Xa21, through particle bombardment. Molecular analysis of transgenic plants revealed the presence of a 3.8-kb EcoRV-digested DNA fragment corresponding to most of the Xa21 coding region and its complete intron sequence, indicating the integration of Xa21 into the genome of ‘IR72’. In the T1 generation, the transgene was inherited and segregated in a 3:1 ratio. After inoculation with the prevalent races 4 and 6 of Xanthomonas oryzae pv. oryzae (Xoo), T1 plants positive for the transgene were found to be resistant to bacterial blight (BB). We also observed that the level of resistance to race 4 of Xoo was higher due to the pyramiding of Xa21 and Xa4 present in ‘IR72’. Since the inactivation of the transgene Xa21 occurred in the two transgenic T1 plants, a larger progeny should be obtained for selecting homozygous line with a consistently higher level of resistance to the BB pathogen.

A benefit of high temperature: increased effectiveness of a rice bacterial blight disease resistance gene.

*Continuous planting of crops containing single disease resistance (R) genes imposes a strong selection for virulence in pathogen populations, often rendering the R gene ineffective. Increasing environmental temperatures may complicate R-gene-mediated disease control because high temperatures often promote disease development and reduce R gene effectiveness. Here, performance of one rice bacterial blight disease R gene was assessed in field and growth chamber studies to determine the influence of temperature on R gene effectiveness and durability. *Disease severity and virulence of Xanthomonas oryzae pv. oryzae (Xoo) populations were monitored in field plots planted to rice with and without the bacterial blight R gene Xa7 over 11 yr. The performance of Xa7 was determined in high- and low-temperature regimes in growth chambers. *Rice with Xa7 exhibited less disease than lines without Xa7 over 11 yr, even though virulence of Xoo field populations increased. Xa7 restricted disease more effectively at high than at low temperatures. Other R genes were less effective at high temperatures. *We propose that Xa7 restricts disease and Xoo population size more efficiently in high temperature cropping seasons compared with cool seasons creating fluctuating selection, thereby positively impacting durability of Xa7.

Mapping quantitative trait loci conferring blast resistance in upland indica rice (Oryza sativa L.)

A genetic analysis of blast resistance in upland rice variety is very crucial. In this study, we performed a linkage mapping of quantitative trait loci (QTLs) for blast resistance using an advanced backcross population from a cross between Way Rarem (susceptible indica variety) and Oryzica Llanos 5 (durable resistant indica variety). A transgressive segregation was observed in the advanced backcross population of Way Rarem//Oryzica Llanos 5. A total of 16 QTLs have been identified along chromosomes 1, 3, 5, 6, 7, 8, 9, and 11 against eight blast pathogen isolates. Each QTL accounted from 11.31 to 45.11% of the variation in blast resistance. Most QTLs showed race specificity, demonstrating the small effect of such QTLs. Unexpectedly, several superior blast resistance alleles were contributed by Way Rarem, the susceptible-recurrent parent. Among eight candidate defense response genes detected in several loci, a single gene (oxalate oxidase) present on chromosome 3 was found to be associated with blast resistance in upland indica rice. Ultimately, these advanced backcross lines with resistance to blast tagged by markers might be useful for pyramiding blast resistance alleles in upland rice.

What it Takes to Achieve Durable Resistance to Rice Blast

Based on performance record, varieties with a reputation for possessing durable blast resistance can be identified from breeding programs. For most of these varieties, a combination of major resistance genes and multiple QTL appears to be the underlying genetic basis for durability. Despite this seemingly simple observation, re-creating genotypes with durable resistance and proper agronomic characteristics remains challenging in breeding. We suggest that genetic mapping, saturation mutagenesis, and transcriptome analysis can together provide a genomewide view of the essential regions for disease resistance, enabling the creation and selection of desirable gene combinations. We further propose actions needed to sustain breeding for durable resistance. These include (a) develop breeding-ready near-isogenic lines for important major resistance (R) genes and QTL, (b) improve knowledge of neck blast through better screening and genetic analysis, (c) promote exchange of breeding materials between countries for evaluation and validation of durable resistance, and (d) resume monitoring of blast pathogen populations with functional diagnostic markers. International collaboration as well as enhanced linkages within countries are much needed to implement these actions.

Chlorophyll Fluorescence and Reflectance-Based Non-Invasive Quantification of Blast, Bacterial Blight and Drought Stresses in Rice

Response of rice (Oryza sativa) exposed to both biotic and abiotic stresses can be quantified by employing fast and accurate optical methods. In this study, the overall stress responses of (i) 12 near-isogenic lines (NILs) in the genetic background of the rice blast-susceptible cultivar Lijiangxintuanheigu (LTH) and (ii) four NILs in the genetic background of the bacterial blight-susceptible cultivar IR24, were inspected by means of Chl fluorescence (Chl-F) imaging. The distribution of the maximum and effective quantum yield of PSII (Fv/FM and QY) and steady-state Chl-F (Ft) were found to be effective in differentiating symptomatic leaf tissue for both rice blast and bacterial blight, which correlated well with 30 cycles of rice blast and six cycles of bacterial blight previously screened using classical (manual) approaches. Subsequently, identified Chl-F parameters allowing detection under ambient light (QY and Ft) were tested across both biotic and abiotic (drought) stress experiments, for rice cultivars contrasting for drought stress response (N22, IR64 and NSIC Rc 222). Their applicability has been proven for both rice blast and bacterial blight; however, QY failed to detect the effect of drought. In addition to Chl-F, the usefulness of 11 selected vegetation indices (Vis) was tested on these three cultivars exposed to particular stresses: (i) rice blast was detectable by Vis calculated from the visible spectrum; (ii) bacterial blight by near-infrared-related Vis; and (iii) drought by Vis calculated from the visible spectrum. The key Chl-F parameters and/or Vis have been summarized and discussed.