Browsing by Author "Lamichhane, Suman"
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- The Central Negative Regulator of Flooding Tolerance, the PROTEOLYSIS 6 Branch of the N-degron Pathway, Adversely Modulates Salinity Tolerance in ArabidopsisLamichhane, Suman; Alpuerto, Jasper B.; Han, Abigail; Fukao, Takeshi (MDPI, 2020-10-23)Seawater intrusion in coastal regions and waterlogging in salinized lands are serious constraints that reduce crop productivity under changing climate scenarios. Under these conditions, plants encounter flooding and salinity concurrently or sequentially. Identification and characterization of genes and pathways associated with both flooding and salinity adaptation are critical steps for the simultaneous improvement of plant tolerance to these stresses. The PROTEOLYSIS 6 (PRT6) branch of the N-degron pathway is a well-characterized process that negatively regulates flooding tolerance in plants. Here, we determined the role of the PRT6/N-degron pathway in salinity tolerance in Arabidopsis. This study demonstrates that the prt6 mutation enhances salinity tolerance at the germination, seedling, and adult plant stages. Maintenance of chlorophyll content and root growth under high salt in the prt6 mutant was linked with the restricted accumulation of sodium ions (Na+) in shoots and roots of the mutant genotype. The prt6 mutation also stimulated mRNA accumulation of key transcription factors in ABA-dependent and independent pathways of osmotic/salinity tolerance, accompanied by the prominent expression of their downstream genes. Furthermore, the prt6 mutant displayed increased sensitivity to ethylene and brassinosteroids, which can suppress Na+ uptake and promote the expression of stress-responsive genes. This study provides genetic evidence that both salinity and flooding tolerance is coordinated through a common regulatory pathway in Arabidopsis.
- Physiological and Molecular Dissection of Salinity Tolerance in Arabidopsis and Maize and Nitrogen Uptake in WheatLamichhane, Suman (Virginia Tech, 2020-04-20)The PROTEOLYSIS 6 (PRT6) branch of the N-end rule pathway is a well-characterized negative regulator of flooding and low oxygen tolerance in plants. This study investigated the role of this pathway in adaptation to salinity stress in Arabidopsis and maize via physiological and molecular characterization of Arabidopsis prt6-1 and maize prt6 MU insertion mutants, respectively. Our study demonstrated that the loss of function mutation of prt6 in Arabidopsis activated hormonal and transcriptional responses associated with adaptation to salinity stress, enhancing high salt tolerance at seed germination, seedling, and adult plant stages. Our data also indicated that salinity tolerance conferred by the prt6 mutation is attributed to increased mRNA abundance of key transcriptional factors in ABA-dependent (AREB/ABFs) and independent (DREBs) pathways, together with the dominant expression of downstream dehydrins. Furthermore, this study revealed that the prt6 mutation enhances ethylene and brassinosteroid responses, resulting in restricted Na+ accumulation in roots and shoots as well as increased expression of dehydrin genes such as RD29A and RD29B. Maize prt6 mutant plants, contrary to our observation in Arabidopsis, showed lower seed germination, primary root elongation, and shoot biomass growth along with increased malondialdehyde (MDA) accumulation under high salt. Moreover, maize prt6 mutants exhibited reduced grain yield and yield-related components under high salt. These results indicate that PRT6 functions as a negative regulator for salinity tolerance in Arabidopsis, whereas this gene plays a positive role in salinity tolerance in maize. In wheat, we compared two genotypes with contrasting nitrogen-use-efficiency (NUE), VA08MAS-369 and VA07W-415, to dissect physiological and molecular mechanisms underlying NUE regulation. Our agronomic data revealed that line 369 maintained yield and yield-related parameters and exhibited greater NUE indexes relative to line 415 under N deficient conditions. Furthermore, our analyses suggested that the significantly higher nitrogen use efficiency (NUE) in line 369 could be attributed to the greater N uptake efficiency in this genotype. In fact, line 369 was able to maintain the development of root systems under N limitation. Consistently, genes encoding high-affinity nitrate transporters such as TaNRT2.1 and TaNRT2.2 were expressed more abundantly in the roots of line 369 than line 415 at limited N. Overall, the results of this study characterized physiological and molecular phenotypes associated with high N uptake efficiency in line 369. This is useful information for the development of new wheat accessions with improved NUE.
- Physiological and Molecular Traits Associated with Nitrogen Uptake under Limited Nitrogen in Soft Red Winter WheatLamichhane, Suman; Murata, Chiaki; Griffey, Carl A.; Thomason, Wade E.; Fukao, Takeshi (MDPI, 2021-01-17)A sufficient nitrogen (N) supply is pivotal for high grain yield and desired grain protein content in wheat (Triticum aestivum L.). Elucidation of physiological and molecular mechanisms underlying nitrogen use efficiency (NUE) will enhance our ability to develop new N-saving varieties in wheat. In this study, we analyzed two soft red winter wheat genotypes, VA08MAS-369 and VA07W-415, with contrasting NUE under limited N. Our previous study demonstrated that higher NUE in VA08MAS-369 resulted from accelerated senescence and N remobilization in flag leaves at low N. The present study revealed that VA08MAS-369 also exhibited higher nitrogen uptake efficiency (NUpE) than VA07W-415 under limited N. VA08MAS-369 consistently maintained root growth parameters such as maximum root depth, total root diameter, total root surface area, and total root volume under N limitation, relative to VA07W-415. Our time-course N content analysis indicated that VA08MAS-369 absorbed N more abundantly than VA07W-415 after the anthesis stage at low N. More efficient N uptake in VA08MAS-369 was associated with the increased expression of genes encoding a two-component high-affinity nitrate transport system, including four NRT2s and three NAR2s, in roots at low N. Altogether, these results demonstrate that VA08MAS-369 can absorb N efficiently even under limited N due to maintained root development and increased function of N uptake. The ability of VA08MAS-369 in N remobilization and uptake suggests that this genotype could be a valuable genetic material for the improvement of NUE in soft red winter wheat.