scholarly journals Aromatic Rice with Multiple Disease Resistance Developed through Anther Culture

2021 ◽  
Vol 12 (5) ◽  
pp. 431-440
Author(s):  
Pawankumar Kharate ◽  
◽  
Pratik Pusadkar ◽  
Nirmala Bharti Patel ◽  
Shrinkhla Maurya ◽  
...  

The experiments were conducted during kharif (June to September) 2018 and 2019 to screen 7 aromatic rice double haploid (DH) lines developed against Bacterial Leaf Bligh (BLB) and blast disease. Three genes Xa21, xa13, xa5 for BLB resistance and two genes Pi1, Pi2 for blast resistance was introgressed into a local aromatic rice variety Dubraj from Chhattisgarh State of India. Dubraj is extremely popular aromatic short medium grain variety in Chhattisgarh and Madhya Pradesh region and hence used in this study. RPBIO-226 and PR122 rice variety is used as donor for BLB and blast resistance respectively. 7 DH lines (L-1 to L-7) were developed from the crosses (Dubraj X RPBIO-226 X PR122) in the year 2017. Phenotypic evaluation for BLB diseases resistance has shown that L-1, L-6 and L-7 (score 1) has a similar resistance as the control check RPBIO-226(score-1) and 3 lines with moderate resistance (L-3 score 2) (L-4 score 3) (L-5 score 3). For blast disease, three lines (L-4, L-6 and L-7) were identified as resistance with the disease score of 1, whereas three were grouped under the section moderately resistant (L-1, L-3 and L- 5) with a disease score of 3. Sensory test (1.7% KOH) was conducted for the estimation of presence / absence of aroma. Moderately strong aroma was present in four lines (L-1, L-3, L-6 and L-7). On the basis of yield and presence of aroma in grain, L-3 has been selected with moderately resistant to BLB and blast for further advancement and subsequently evaluated under State Initial Evaluation Trial (SIET) during kharif 2020. Within 3 years of development of DH line, L-3 is under varietal developmental trail, which itself indicate the efficacy of anther culture in fixing homozygosity and speedy development of desired variety within short period of time.

2021 ◽  
Author(s):  
◽  
Fabiano Touzdjian Pinheiro Kohlrausch Távora

Rice (Oryza sativa L.) is the main food crop for more than half of the world population but unfortunately, it is severely affected by blast, one of the most widespread and devastating plant diseases, caused by the fungus Magnaporthe oryzae. Hence, the development of rice cultivars with greater resistance to blast is one of the main focuses of breeding programs. However, due to the complex biology of the pathogen, rice cultivars genetically resistant to the fungus become susceptible in a short period of time. In this context, the knockout of rice susceptibility genes represents a flourishing approach to obtain rice cultivars with a broader and longer-lasting resistance to M. oryzae. The present study aimed to use the genomic editing technology - CRISPR/Cas9 system, for knocking-out genes engaged with rice susceptibility to fungal infection. From previous transcriptomics results of two semi-isogenic rice lines - NILs infected by M. oryzae, potential rice-blast susceptibility genes were selected. The prospection of candidate genes for gene editing was complemented by a comparative shotgun proteomic analysis of the protein profile of the interaction between IRBLi-F5 (susceptible) and IRBL5-M (resistant) NILs in early stages of M. oryzae infection, that revealed a specific set of proteins potentially associated with susceptibility. After the characterization and validation of gene expression by RT-qPCR of the most prominent candidates, the target genes OsDjA2, OsERF104 and OsPyl5 were selected and submitted to a functional validation via gene silencing in planta, using antisense oligonucleotides (ASO), in which a clear reduction of leaf symptoms was observed in the compatible identification. Subsequently, the model japonica rice variety Nipponbare was transformed with simplex CRISPR/Cas9 vectors aiming to the independent knockout of each target gene. The T1 progeny of rice-edited plants, homozygous for the null (loss of function)-mutation were tested for blast resistance. As expected, mutant plants showed a decrease of disease symptoms in comparison with control lines (transformant non-edited plants). The results obtained in this study can contribute for the development of rice cultivars resistant to blast disease, besides shedding light on new potential rice-blast susceptibility genes.


2019 ◽  
Vol 2019 ◽  
pp. 1-10
Author(s):  
L. Kanyange ◽  
J. Kamau ◽  
O. Ombori ◽  
A. Ndayiragije ◽  
M. Muthini

The ascomycete fungus, Pyricularia oryzae or Magnaporthe oryzae, is known to cause blast disease in more than 80 host plants of the Gramineae family—cereals including rice and grasses. The improvement of the Supa234 rice line (IR97012-27-3-1-1-B, containing badh2 gene for aroma) developed at IRRI-ESA Burundi consisted of introgression of R genes (Pita and Pi9) for blast resistance. The F2 population obtained via the cross had been screened for blast resistance using inoculation with Pyricularia oryzae spore’s suspension. The objectives of this study were to assess the presence of Pita and Pi9 genes for blast resistance and to assess the presence of the badh2 gene for aroma in the screened F2 plants using molecular markers. Genotyping was carried out in 103 F2 plants which grew to maturity using the KASP genotyping method with SNP markers (snpOS0007, snpOS0006, and snpOS0022) targeting the Pita and Pi9 genes for blast resistance and the badh2 gene for aromatic fragrance. The genotyping results showed that 38 F2 plants had the Pita gene present in both alleles, 31 F2 plants with the Pita gene in one allele, and only one plant (3B1) was found with the Pi9 gene in one allele. The badh2 gene for aroma was detected in 27 F2 plants on both alleles and in 57 F2 plants on one allele. There were thirteen plants which had both the Pita gene and the badh2 gene for aroma, and only one plant (3B1) had a combination of the three genes (Pita, Pi9, and badh2). Seven plants resistant to blast disease (2H2, 2H4, 1G2, 1C12, 1E13, 1B12, and 1C5) with the Pita and badh2 genes were found, and only one resistant plant (3B1) had a combination of the three genes Pi9, Pita, and badh2 which is recommended to be bulked for the development of the Supa aromatic rice variety resistant to blast disease. The plants generated by the best line 3B1 should further be evaluated for grain quality (Supa type) after F5 generation in the field.


2021 ◽  
Author(s):  
Xingyu An ◽  
Hui Zhang ◽  
Jinlu Li ◽  
Rui Yang ◽  
Qianchun Zeng ◽  
...  

Abstract Background: The molecular mechanism of durable and broad-spectrum resistance to rice blast disease in japonica rice variety is still very little known. Ziyu44, a local japonica rice variety in Yunnan Province of China, has shown durable broad-spectrum blast resistance for more than 30 years, and provides an opportunity for us to explore the molecular basis of broad-spectrum resistance to rice blast in japonica rice variety.Methods and Results: We conducted a comparative study of mycelium growth, aposporium formation, the accumulation of salicylate(SA), jasmonate(JA) and H2O2, the expression of SA- and JA-associated genes between Ziyu44 and susceptible variety Jiangnanxiangnuo (JNXN) upon M. oryzae infection. We found that appressorium formation and invasive hyphae extention were greatly inhibited in Ziyu 44 leaves compared with that in JNXN leaves. Both Ziyu 44 and JNXN plants maintained high levels of baseline SA and did not show increased accumulation of SA after inoculation with M. oryzae, while the levels of baseline JA in Ziyu 44 and JNXN plants were relatively low, and the accumulation of JA exhibited markedly increased in Ziyu 44 plants upon M. oryzae infection. The expression levels of key genes involving JA and SA signaling pathway OsCOI1b, OsNPR1, OsMPK6 as well as pathogenesis-related (PR) genes OsPR1a, OsPR1b and OsPBZ1, were markedly up-regulated in Ziyu44. Conclusions: The level of endogenous JA is critical for synchronous activation of SA and JA signaling pathway, up-regulating PR gene expression and enhancing disease resistance against rice blast in Ziyu44.


Author(s):  
KD Puri ◽  
SM Shrestha ◽  
KD Joshi ◽  
GB KC

The severity of the rice blast disease (Pyricularia grisea) of both leaf and neck varies with different environment and it becomes destructive under favorable condition. The leaf and neck blast resistance and susceptible interaction of 30 different tropical rice lines were evaluated under low-, mid- and up-land conditions of Chitwan district and classified on the basis of disease severity with respect to susceptible check, Masuli. Of them, 5, 10, 12 and 3 rice lines were resistant to leaf blast, moderately resistant, moderately susceptible susceptible, respectively. Similarly, for the neck blast nine lines were resistant, thirteen moderately resistant, seven moderately susceptible and one was susceptible. The progenies from Masuli/MT4 had the highest leaf and neck blast susceptible reaction, while the most of progenies from IPB (Irradiated Pusa Basmati), KalinghaIII_IR64, Radha 32_ KIII and Masuli_IR64 were resistant, and the most promising sources against leaf and neck blast resistance. Therefore, the progenies from these parents can be used in breeding the resistant variety. Key words: Pyricularia grisea, resistance, rice lines J. Inst. Agric. Anim. Sci. 27:37-44 (2006)


Agronomy ◽  
2019 ◽  
Vol 9 (2) ◽  
pp. 67 ◽  
Author(s):  
Nawaporn Srichant ◽  
Sompong Chankaew ◽  
Tidarat Monkham ◽  
Petcharat Thammabenjapone ◽  
Jirawat Sanitchon

The Sakon Nakhon (SKN) rice cultivar is non-photosensitive, rich in aroma, and an excellent choice for cooking quality; which adapts well to both upland and lowland conditions. Furthermore, it has been adopted by Thailand’s sugarcane growers, as it has proved fit for an upland crop preceding the sugarcane cropping system. However, SKN is very susceptible to blast disease caused by Pyricularia oryzae. The purpose of this study was to breed SKN for blast resistance, using marker-assisted backcrossing, together with phenotypic selection. The breeding population was constructed by crossing the SKN (recurrent) with the RD6 introgression line (donor), which contained four blast resistant quantitative trait loci (QTLs). Franking microsatellite markers for blast resistance QTLs on chromosomes 1, 2, 11 and 12 were used for selection through BC2F2. Thirteen BC2F3 selected lines were evaluated under greenhouse conditions, and seven lines of BC2F4 were evaluated against blast, along with their agronomic traits, under field conditions (upland and lowland). SKN 39-10-19-29-12 and SKN 39-10-19-29-13 presented high resistance to leaf and neck blast; and were non-photosensitive, with the same agronomic traits as the SKN. This study successfully demonstrates the improvement of the recommended SKN variety blast resistance, while maintaining similar agronomic traits.


2021 ◽  
Vol 32 (3) ◽  
pp. 147-160
Author(s):  
Siti Norsuha Misman ◽  
Mohd Shahril Firdaus Ab Razak ◽  
Nur Syahirah Ahmad Sobri ◽  
Latiffah Zakaria

Rice blast caused by Pyricularia oryzae (P. oryzae) is one of the most serious diseases infecting rice worldwide. In the present study, virulence pattern of six P. oryzae pathotypes (P0.0, P0.2, P1.0, P3.0, P7.0 and P9.0) identified from the blast pathogen collected in Peninsular Malaysia, were evaluated using a set of 22 IRRI-bred blast resistance lines (IRBL) as well as to determine the resistance genes involved. The information on the virulence of the blast pathotypes and the resistance genes involved is important for breeding of new rice variety for durable resistance against blast disease. The IRBL was established from 22 monogenic lines, harbouring 22 resistance genes [Pia, Pib, Pii, Pit, Pi3, Pi5(t), Pish, Pi1, Pik, Pik-s, Pik-m, Pik-h, Pik-p, Pi7(t), Pi9, Piz, Piz-5, Piz-t, Pi19, Pi20(t), Pita-2, and Pita=Pi4(t)]. Based on the disease severity patterns, the tested pathotypes were avirulence towards seven IRBLs [IRBLi-F5, IRBLk-Ka, IRBLkh-K3, IRBLz-Fu, IRBLsh-S, IRBLPi7 (t) and IRBL9-W] of which these IRBLs harbouring Pii, Pik, Pik-h, Piz, Pish, Pi7(t) and Pi9 resistance genes, respectively. Therefore, the results suggested that the seven IRBLs carrying seven resistance genes [Pii, Pik, Pik-h, Piz, Pish, Pi7(t) and Pi9] would be suitable candidates of resistance genes to be incorporated in new breeding lines to combat the current blast pathotypes in the field.


Plant Disease ◽  
2021 ◽  
Author(s):  
Yulin Jia ◽  
Melissa H Jia ◽  
Zongbu Yan

Rice blast disease caused by the fungus Magnaporthe oryzae (syn. M. grisea) is one of the most lethal diseases for sustainable rice production worldwide. Blast resistance mediated by major resistance genes are often broken-down after a short period of deployment, while minor blast resistance genes, each providing a small effect on disease reactions, are more durable. In the present study, we first evaluated disease reactions of two rice breeding parents ‘Minghui 63’ and ‘M-202’ with 11 US blast races, IA45, IB1, IB45, IB49, IB54, IC1, IC17, ID1, IE1, IG1, and IH1 commonly found under greenhouse conditions using a category disease rating resembling infection types under field conditions. ‘Minghui 63’ exhibited differential resistance responses in comparison with that of ‘M-202’ to the tested blast races. A recombinant inbred line (RIL) population of 275 lines from a cross between ‘Minghui 63’ and ‘M-202’ was also evaluated with the above mentioned blast races. The population was genotyped with 156 simple sequence repeat (SSR) and insertion and deletion (Indel) markers. A linkage map with a genetic distance of 1022.84 cM was constructed using inclusive composite interval mapping (ICIM) software. A total of 10 resistance QTLs, eight from ‘Minghui 63’ and two from ‘M-202’, were identified. One major QTL, qBLAST2 on chr 2, was identified by seven races/isolates. The remaining nine minor resistance QTLs were mapped on chromosome 1, 3, 6, 9, 10, 11 and 12. These findings provide useful genetic markers and resources to tag minor blast resistance genes for marker assisted selection in rice breeding program and for further studies of underlying genes.


2020 ◽  
Vol 56 (No. 3) ◽  
pp. 93-101
Author(s):  
Bo Lan ◽  
Ying-Qing Yang ◽  
Qiang Sun ◽  
Hong-Fan Chen ◽  
Jian Chen ◽  
...  

To understand the cause of loss of rice blast resistance, we studied the pathogenicity of Magnaporthe oryzae strains isolated from rice hybrid Wuyou 308 and evaluated its resistance genes. A total of 62 M. oryzae strains were isolated and tested in 7 Chinese rice varieties with varying degrees of resistance to rice blast and 30 blast-resistant monogenic lines. Fourteen physiological races of M. oryzae were identified: 8.55% belonging to the ZA group, 86.67% to the ZB group, and 5.00% to the ZC group. ZB15 was the most abundant race (45.00%). Five resistance genes, Pi-3(1), Pi-z5, Pi-k, Pi-kp(C), and Pi-k(C), conferred good resistance to the 62 strains, with resistance frequencies of 95.56, 91.11, 88.89, 82.22, and 82.22%, respectively. In contrast, Pi-a(2) had a resistance frequency of 0%. The hybrid combination Wuyou 308 was found to carry Pi-ta and Pi-b genes. Because Pi-ta and Pi-b both showed low resistance frequencies to M. oryzae isolated from Jiangxi, the hybrid rice variety Wuyou 308 could be infected by most of the 62 M. oryzae strains. The emergence and spread of rice blast disease in Wuyou 308 may thus be difficult to avoid when climatic conditions are favourable.


2020 ◽  
Vol 3 (2) ◽  
pp. 276-286
Author(s):  
Pratiksha Sharma ◽  
Prem Bahadur Magar ◽  
Suraj Baidya ◽  
Ram Baran Yadaw

Blast, caused by Pyriculariagrisea (Sacc.) is the most destructive disease of rice in Nepal. To identify the sources of leaf blast resistance in rice genotypes, a field experiment was conducted under natural epiphytotic condition at National Plant Pathology Research Centre (NPPRC), Khumaltar, Lalitpur, Nepal during summer season in 2018 and 2019.A total of 128 rice genotypes in 2018 and 291during 2019 including resistant check (Sabitri) and susceptible check (Shankharika/Mansuli) were tested. Field experiment was conducted in single rod row design. Leaf blast disease assessment was done according to 0-9 scale. During 2018, 59 entries were highly resistant (Score 0), 34 resistant (Score 1), 26 moderately resistant (Score 2-3), 5 were moderately susceptible (Score 4-5), 4 susceptible (Score 6-7) and none of them were highly susceptible (Score 8-9) to leaf blast. Similarly, in 2019, 6 lines were highly resistant, 70 resistant, 196 moderately resistant, 15 lines were moderately susceptible, 4 susceptible and none of them were highly susceptible to the disease. Only, one genotype NR2179-82-2-4-1-1-1-1 (Score 1) was found resistant in both years. Similarly, genotype NR2182-22-1-3-1-1-1 (Score 2-3) was found moderately resistant. Some of the genotypes were found resistant in 2018 which become moderately resistant in 2019, they were NR2180-20-2-5-1-1-1-1, IR97135-8-3-1-3, IR98786-13-1-2-1, NR2181-139-1-3-1-1-1-1, and IR13F402. So, findings of these resistant and moderately resistant genotypes could be used in resistant source for the development of leaf blast resistant rice varieties through hybridization in future.


2017 ◽  
Vol 9 (4) ◽  
pp. 371-377
Author(s):  
GUT WINDARSIH ◽  
DWINITA WIKAN UTAMI

Windarsih G, Utami DW. 2017. Evaluation of neck blast resistance and agronomical performances on double haploid rice population in greenhouse and endemic field. Nusantara Bioscience 9: 371-377. Blast disease caused by fungal Pyricularia grisea Sacc. is one of the most destructive diseases of rice in the world. The development of blast-resistant rice varieties will be essential to control this disease. This research aimed (i) to compare the resistance response to neck-blast among DH lines from double cross IR54/Parekaligolara//Bio110/Markuti and the differential varieties against three selected Indonesian blast races in greenhouse, (ii) to identify the gene(s) that caused the resistance to neck-blast based on the association between the resistance response and the genotype evaluation using molecular markers linked to Pi1, Pi33, Pib, Pir4 and Pir7 genes, and (iii) to evaluate the resistance response to leaf and neck blast on DH lines in endemic field (Sukabumi) and the agronomical performance of selected DH lines in optimum field in Ciasem of Subang, West Java, Indonesia during December 2013 to March 2014. Eleven double haploid lines from double-crossing IR54/Parekaligolara//Bio110/Markuti, the differential varieties as resistant control and the US2 variety for susceptible control were observed for neck-blast resistance response to three blast races in greenhouse and endemic field (Sukabumi), while the agronomical performances were observed in field of Ciasem-Subang. The results based on the genotyping evaluation, leaf and neck blast resistance, either in greenhouse and endemic location, and the agronomical performance in field showed that 5 selected double haploid lines had leaf and neck blast resistance and good performance on field trial. Thus they are promising for use either for further testing forwarding into releasing variety or used as donor for further blast resistant breeding activities.


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