Abiotic stresses, such as salt and heat stress, coexist in some regions of the world and can have a significant impact on agricultural plant biomass and production. Rice is a valuable crop that is susceptible to salt and high temperatures. Here, we studied the role of flavanol 3-hydroxylase in response to combined salt and heat stress with the aim of better understanding the defensive mechanism of rice. We found that, compared with wild-type plants, the growth and development of transgenic plants were improved due to higher biosynthesis of kaempferol and quercetin. Furthermore, we observed that oxidative stress was decreased in transgenic plants compared with that in wild-type plants due to the reactive oxygen species scavenging activity of kaempferol and quercetin as well as the modulation of glutathione peroxidase and lipid peroxidase activity. The expression of high-affinity potassium transporter (HKT) and salt overly sensitive (SOS) genes was significantly increased in transgenic plants compared with in control plants after 12 and 24 h, whereas sodium-hydrogen exchanger (NHX) gene expression was significantly reduced in transgenic plants compared with in control plants. The expression of heat stress transcription factors (HSFs) and heat shock proteins (HSPs) in the transgenic line increased significantly after 6 and 12 h, although our understanding of the mechanisms by which the F3H gene regulates HKT, SOS, NHX, HSF, and HSP genes is limited. In addition, transgenic plants showed higher levels of abscisic acid (ABA) and lower levels of salicylic acid (SA) than were found in control plants. However, antagonistic cross talk was identified between these hormones when the duration of stress increased; SA accumulation increased, whereas ABA levels decreased. Although transgenic lines showed significantly increased Na+ ion accumulation, K+ ion accumulation was similar in transgenic and control plants, suggesting that increased flavonoid accumulation is crucial for balancing Na+/K+ ions. Overall, this study suggests that flavonoid accumulation increases the tolerance of rice plants to combined salt and heat stress by regulating physiological, biochemical, and molecular mechanisms.
Efficient accumulation of flavonoids is important for increased tolerance to biotic stress. Although several plant defense mechanisms are known, the roles of many pathways, proteins, and secondary metabolites in stress tolerance are unknown. We generated a flavanone 3-hydroxylase (F3H) overexpressor rice line and inoculated Xanthomonas Oryzae pv. oryzae and compared the control and wildtype inoculated plants. In addition to promoting plant growth and developmental maintenance, the overexpression of F3H increased the accumulation of flavonoids and increased tolerance to bacterial leaf blight (BLB) stress. Moreover, leaf lesion length was higher in the infected wildtype plants compared with infected transgenics. Kaempferol and quercetin, which scavenge reactive oxygen species, overaccumulated in transgenic lines compared with wildtypes in response to pathogenic infection, detected by scanning electron microscopy and spectrophotometry. The induction of F3H altered the antioxidant system and reduced the levels of glutathione peroxidase activity and malondialdehyde (MDA) contents in the transgenic lines compared with the wildtypes. Downstream gene regulation analysis showed that the expression of F3H increased the regulation of flavonol synthase (FLS), dihydroflavonol 4-reductase (DFR), and slender rice mutant (SLR1) during BLB stress. The analysis of SA and JA signaling revealed an antagonistic interaction between both hormones and that F3H induction significantly promoted SA and inhibited JA accumulation in the transgenic lines. SA-dependent nonexpressor pathogenesis-related (NPR1) and Xa1 showed significant upregulation in the infected transgenic lines compared with the infected control and wildtype lines. Thus, the overexpression of F3H was essential for increasing BLB stress tolerance.
Soil salinity is a major problem in agriculture because high accumulation of Na+ ions in plants causes toxicity that can result in yield reduction. Na+/K+ homeostasis is known to be important for salt tolerance in plants. Na+/K+ homeostasis in rice (Oryza sativa L.) involves nine high-affinity K+ transporter (HKT) encoding Na+-K+ symporter, five OsNHX Na+/H+ antiporters, and OsSOS1 Na+/K+ antiporter genes. In the present study, we investigated various molecular and physiological processes to evaluate germination rate, growth pattern, ion content, and expression of OsHKT, OsNHX, and OsSOS1genes related to Na+/K+ homeostasis in different rice genotypes under salt stress. We found a significant increase in the germination percentage, plant vigor, Na+/K+ ratio, and gene expression of the OsHKT family in both the roots and shoots of the Nagdong cultivar and salt-tolerant cultivar Pokkali. In the roots of Cheongcheong and IR28 cultivars, Na+ ion concentrations were found to be higher than K+ ion concentrations. Similarly, high expression levels of OsHKT1, OsHKT3, and OsHKT6 were observed in Cheongcheong, whereas expression levels of OsHKT9 was high in IR28. The expression patterns of OsNHX and OsSOS1 and regulation of other micronutrients differed in the roots and shoots regions of rice and were generally increased by salt stress. The OsNHX family was also expressed at high levels in the roots of Nagdong and in the roots and shoots of Pokkali; in contrast, comparatively low expression levels were observed in the roots and shoots of Cheongcheong and IR28 (with the exception of high OsNHX1 expression in the roots of IR28). Furthermore, the OsSOS1 gene was highly expressed in the roots of Nagdong and shoots of Cheongcheong. We also observed that salt stress decreases chlorophyll content in IR28 and Pokkali but not in Cheongcheong and Nagdong. This study suggests that under salt stress, cultivar Nagdong has more salt-tolerance than cultivar Cheongcheong.
Drought and ultraviolet radiation (UV radiation) are the coexisting environmental factors that negatively affect plant growth and development via oxidative damage. Flavonoids are reactive, scavenging oxygen species (ROS) and UV radiation-absorbing compounds generated under stress conditions. We investigated the biosynthesis of kaempferol and quercetin in wild and flavanone 3-hydroxylase (F3H) overexpresser rice plants when drought and UV radiation stress were imposed individually and together. Phenotypic variation indicated that both kinds of stress highly reduced rice plant growth parameters in wild plants as compared to transgenic plants. When combined, the stressors adversely affected rice plant growth parameters more than when they were imposed individually. Overaccumulation of kaempferol and quercetin in transgenic plants demonstrated that both flavonoids were crucial for enhanced tolerance to such stresses. Oxidative activity assays showed that kaempferol and quercetin overaccumulation with strong non-enzymatic antioxidant activity mitigated the accumulation of ROS under drought and UV radiation stress. Lower contents of salicylic acid (SA) in transgenic plants indicated that flavonoid accumulation reduced stress, which led to the accumulation of low levels of SA. Transcriptional regulation of the dehydrin (DHN) and ultraviolet-B resistance 8 (UVR8) genes showed significant increases in transgenic plants compared to wild plants under stress. Taken together, these results confirm the usefulness of kaempferol and quercetin in enhancing tolerance to both drought and UV radiation stress.
Rice is one of the world’s leading food crops, and over 90% of the world’s rice production stems from Asia. In particular, an increase of 1 °C in the minimum temperature reduces the quantity of rice by 10%. Therefore, the development of rice varieties that can stably maintain the yield and quality of the rice even under these rapid climate changes is indispensable. In this study, we performed quantitative trait loci (QTL) mapping after treatment with heat stress during the booting stage in rice. We performed a QTL analysis using the Cheongcheong/Nagdong double haploid (CNDH) line and identified 19 QTLs during the 2 year analysis. Of these QTL regions, the 2.2 cM region of RM3709–RM11694 on chromosome 1 was shared among the six traits (heading date; culm length; panicle length; number of tiller; 1000 grain weight; and content of chlorophyll) examined. Rice Microsatellite (RM) 3709–RM11694 contained 27 high-temperature-tolerance candidate genes. Among the candidate genes, OsBHT showed a different gene expression level between CNDH75, which is a high-temperature tolerant line, and CNDH11 which is a susceptible line. Although some existing high-temperature-tolerant genes have been reported, OsBHT can be used more effectively for the development of heat tolerance in rice.
The assessment of the genetic diversity within germplasm collections can be accomplished using simple sequence repeat (SSR) markers and association mapping techniques. The present study was conducted to evaluate the genetic diversity of a colored rice germplasm collection containing 376 black-purple rice samples and 172 red pericarp samples, conserved by Dong-A University. There were 600 pairs of SSR primers screened against 11 rice varieties. Sixteen informative primer pairs were selected, having high polymorphism information content (PIC) values, which were then used to assess the genetic diversity within the collection. A total of 409 polymorphic amplified fragments were obtained using the 16 SSR markers. The number of alleles per locus ranged from 11 to 47, with an average of 25.6. The average PIC value was 0.913, ranging from 0.855 to 0.964. Four hundred and nine SSR loci were used to calculate Jaccard’s distance coefficients, using the unweighted pair-group method with arithmetic mean cluster analysis. These accessions were separated into several distinctive groups corresponding to their morphology. The results provided valuable information for the colored rice breeding program and showed the importance of protecting germplasm resources and the molecular markers that can be derived from them.
Whitebacked planthopper (WBPH) is a pest that causes serious damage to rice in Asian countries with a mild climate. WBPH causes severely rice yield losses and grain poor quality each year so needs biological control. Plants resist biotic and abiotic stress using expressing variety genes, such as kinase, phytohormones, transcription factors, and especially secondary metabolites. In this research, quantitative trait locus (QTL) mapping was performed by assigning the WBPH resistance score in the Cheongcheong/Nagdong doubled haploid (CNDH) line in 2018 and 2019. The RM280-RM6909 on chromosome 4 was detected as a duplicate in 2018, 2019, and derived from Cheongcheong. This region includes cell function, kinase, signaling, transcription factors, and secondary metabolites that protect plants from the stress of WBPH. The RM280-RM6909 on chromosome 4 contains candidate genes that are similar to the flavanone 3-hydroxylase (F3H) of rice. The F3H are homologous genes, which play an important role in biosynthesis defending against biotic stress in plants. After WBPH inoculation, the relative expression level of F3H was higher in resistant line than in a susceptible line. The newly identified WBPH resistance gene F3H by QTL mapping can be used for the breeding of rice cultivars that are resistant against WBPH.
Background Recent temperature increases due to rapid climate change have negatively affected rice yield and grain quality. Particularly, high temperatures during right after the flowering stage reduce spikelet fertility, while interfering with sugar energy transport, and cause severe damage to grain quality by forming chalkiness grains. The effect of high-temperature on spikelet fertility and grain quality during grain filling stage was evaluated using a double haploid line derived from another culture of F1 by crossing Cheongcheong and Nagdong cultivars. Quantitative trait locus (QTL) mapping identifies candidate genes significantly associated with spikelet fertility and grain quality at high temperatures. Results Our analysis screened OsSFq3 that contributes to spikelet fertility and grain quality at high-temperature. OsSFq3 was fine-mapped in the region RM15749-RM15689 on chromosome 3, wherein four candidate genes related to the synthesis and decomposition of amylose, a starch component, were predicted. Four major candidate genes, including OsSFq3, and 10 different genes involved in the synthesis and decomposition of amylose and amylopectin, which are starch constituents, together with relative expression levels were analyzed. OsSFq3 was highly expressed during the initial stage of high-temperature treatment. It exhibited high homology with FLOURY ENDOSPERM 6 in Gramineae plants and is therefore expected to function similarly. Conclusion The QTL, major candidate genes, and OsSFq3 identified herein could be effectively used in breeding rice varieties to improve grain quality, while tolerating high temperatures, to cope with climate changes. Furthermore, linked markers can aid in marker-assisted selection of high-quality and -yield rice varieties tolerant to high temperatures.
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