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Physiological and Molecular Dissection of Salinity Tolerance in Arabidopsis and Maize and Nitrogen Uptake in WheatLamichhane, Suman 20 April 2020 (has links)
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. / Doctor of Philosophy / In coastal areas, sea-level rise increases the chances of saltwater intrusion into cultivable lands, making a hostile environment for crop growth and production by imposing flooding and salinity stresses simultaneously. Identification of central regulators that regulate the adaptation to both flooding and salinity is a critical step for the development of new crop genotypes with enhanced tolerance to these stresses. Previous studies have characterized the function of the PROTEOLYSIS 6 (PRT6) gene in adaptation to flooding stress in plants. This study assessed whether this gene is involved in adaptation to salinity stress in Arabidopsis and maize by evaluating the growth and survival of their respective prt6 mutants under high salt. Consistent with the flooding tolerance data, our study showed that the PRT6 gene also functions as a negative regulator of salinity stress tolerance in Arabidopsis. The prt6 mutation in Arabidopsis activated the key transcriptional and hormone response pathways associated with adaptation to both salinity/osmotic stress and sodium toxicity, expressed as enhanced tolerance to excess salt at seed germination, seedling, and adult plant stages. In maize, disruption of the PRT6 gene decreased seed germination, primary root elongation, and shoot biomass growth under high salt, which is opposite to our observations in Arabidopsis. Additionally, the maize mutant plants encountered more oxidative stress, as demonstrated by the higher accumulation of malondialdehyde (MDA) under high salt. Moreover, maize prt6 mutants exhibited reduced grain yield under high salt. Overall, these results indicate that disruption of the PRT6 gene confers increased tolerance to high salt in Arabidopsis, whereas it conversely reduced salinity tolerance in maize. In wheat, we compared two genotypes with distinct nitrogen use efficiency (NUE), VA08MAS-369 and VA07W-415, to determine critical traits involved in NUE regulation. Our study showed that grain yield and yield-related parameters were significantly higher in line 369 than line 415 under low N. Moreover, high NUE in line 369 was attributed to efficient N uptake in this genotype under limited N. Our root architecture analysis demonstrated that line 369 was able to maintain root depth, volume, and thickness even under N limitation. Consistently, line 369 highly induced expression of genes associated with nitrogen transport at low N. Altogether, this study identified key traits involved in high NUE in wheat, facilitating the breeding of new wheat genotypes with enhanced NUE.
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MEASURING SOIL NITROUS OXIDE EMISSIONS BY USING A NOVEL OPEN PATH SCANNING TECHNIQUECheng-Hsien Lin (5929973) 02 August 2019 (has links)
A
better way to improve understanding and quantification of nitrous oxide (N<sub>2</sub>O)
emitted from intensive maize cropping systems is to develop an advanced emissions
measurement method This study developed an open path (OP) method to measure N<sub>2</sub>O
emissions from four adjacent maize plots managed by tillage practices of no-till
(NT) and chisel plow (ChP), and different nitrogen (N) treatments from 2014 to
2016. Anhydrous ammonia (220 kg NH<sub>3</sub>-N ha<sup>-1</sup>) was applied in once or equally split (full vs.
split rate) and applied in different timing (Fall vs. Spring). The spring N
application occurred either before planting (pre-plant) or in season (side-dress).
Emissions measurements were conducted by using
the OP method (the scanning OP Fourier transform infrared spectrometry (OP-FTIR)
+ the gas point-sampling system + a backward Lagrangian stochastic (bLS)
dispersion model) and static closed chamber methods. The performance and
feasibility of the OP measurements were
assessed by a sensitivity analysis, starting with errors associated with the
OP-FTIR for calculating N<sub>2</sub>O concentrations, and then errors
associated with the bLS model for
estimating N<sub>2</sub>O emissions. The quantification of N<sub>2</sub>O
concentrations using the OP-FTIR spectrum was influenced by ambient humidity,
temperature, and the path length between a spectrometer and a retro-reflector.
The optimal quantitative method mitigated these ambient interference effects on
N<sub>2</sub>O quantification. The averaged bias of the calculated N<sub>2</sub>O
concentrations from the spectra acquired from wide ranges of humidity (0.5 – 2.0
% water vapor content), temperature (10 – 35 °C), and path length (100 – 135
meters) was 1.4 %. The precision of the OP-FTIR N<sub>2</sub>O concentrations
was 5.4 part
per billion<sup> </sup>(3σ) in a stationary flow condition for a 30-minute averaging period. The emissions
measurement from multiple sources showed that the field of interest was likely
interfered by adjacent fields. Fields with low emission rates were more sensitive
to the adjacent fields with high emissions, resulting in substantial biases and
uncertainties. The minimum detection limit of the N<sub>2</sub>O emission rates
was 1.2 µg m<sup>-2</sup> s<sup>-1</sup> (MDL; 3σ). The OP measurements showed
that the NT practice potentially reduced N<sub>2</sub>O emission compared with ChP. Under the long-term NT treatments, the
split-N rate application (110 kg NH<sub>3</sub>-N ha<sup>-1</sup> in the fall
and spring) resulted in lower N<sub>2</sub>O emissions than the full
application (220 kg NH<sub>3</sub>-N ha<sup>-1</sup> in the fall). The management
of NT coupled with split-N rate application minimized N<sub>2</sub>O emissions among
treatments in this study, resulting in N<sub>2</sub>O-N losses of 3.8, 13.2,
and 6.6 N kg ha<sup>-1</sup> over 9-, 35-, and 20-days after the spring NH<sub>3</sub>
application in 2014, 2015, and 2016, respectively. The spring pre-plant N
application in 2015 also resulted in higher N<sub>2</sub>O emissions than the
spring side-dress application in 2016, and the increased N<sub>2</sub>O-N loss
was corresponding to lower N recovery efficiency in 2015 measurements. A
comparison of chamber and OP measurements showed that soil N<sub>2</sub>O
emissions were likely underestimated by 10x without considering the
wind-induced effect on gas transport at the ground-atmospheric interface. This
study showed that the OP method provides a great
opportunity to study agricultural N<sub>2</sub>O emissions as well as management optimization for the sustainability
of the agroecosystems.
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Evaluation of Eggplant, Wild Relatives and Introgression Breeding Materials for Tolerance to Abiotic StressesVillanueva Párraga, Gloria 07 October 2024 (has links)
Tesis por compendio / [ES] El cambio climático, impulsado por actividades humanas, intensifica el estrés abiótico en la agricultura, afectando la productividad y la salud del ecosistema. La creciente demanda de productos vegetales expande las tierras agrícolas, empeorando el cambio climático y la desertificación. El estrés por sequía afecta gravemente la productividad de los cultivos, mientras que la fertilización excesiva con nitrógeno altera el ciclo del nitrógeno, causando problemas ambientales y de salud.
La berenjena (Solanum melongena L.) es una hortaliza importante, y el uso de recursos genéticos, incluidos los parientes silvestres, es crucial para los programas que buscan mejorar la tolerancia a la sequía y la eficiencia en el uso del nitrógeno. Los avances en secuenciación y genotipado han mejorado nuestra comprensión de los rasgos relacionados con el estrés en los cultivos.
Esta tesis doctoral explora el potencial de los parientes silvestres de la berenjena para mejorar la tolerancia a los estreses abióticos. El objetivo es identificar nuevas variaciones genéticas para programas de mejora, centrándose en recursos genéticos, regiones genómicas y genes candidatos para obtener variedades de berenjena más resilientes mediante análisis genómicos, transcriptómicos y fenotípicos.
El capítulo I evalúa retrocruzamientos avanzados (ABs) de berenjena y parientes silvestres para mejorar la eficiencia en el uso del nitrógeno (NUE) en condiciones de bajo nitrógeno (N). El estudio de S. elaeagnifolium y los ABs en el fondo genético de la berenjena en condiciones de bajo N reveló diferencias significativas entre los parentales. La alta diversidad fenotípica en los ABs mostró algunos individuos transgresivos con mayores rendimientos y NUE que el parental cultivado. Las evaluaciones de ABs de diferentes bancos genéticos (S. insanum, S. dasyphyllum y S. elaeagnifolium) en condiciones de bajo N destacaron el potencial de estas especies silvestres. La alta diversidad en los ABs respalda el potencial de los parientes silvestres para mejorar la resiliencia de la berenjena bajo estrés abiótico. Los datos de genotipado y fenotipado permitieron la identificación y validación de QTLs para varios rasgos, ampliando la comprensión genómica de la berenjena.
El capítulo II investiga los mecanismos de respuesta a la sequía en la berenjena cultivada (S. melongena) y su pariente silvestre S. dasyphyllum. Evaluados en condiciones hidropónicas con concentraciones de polietilenglicol (PEG) en diferentes etapas fenológicas, la secuenciación de ARN (RNA-Seq) analizó los patrones de expresión génica. S. dasyphyllum demostró una tolerancia superior al estrés osmótico en comparación con S. melongena, con más genes diferencialmente expresados (DEGs) bajo estrés. Los análisis de enriquecimiento GO y las vías KEGG revelaron que ambas especies activaron varios factores de transcripción y vías de respuesta al estrés, con S. dasyphyllum mostrando una regulación génica más extensa. Las vías clave incluyeron la señalización de ABA, la señalización de MAPK y varias vías de biosíntesis. Estos resultados destacan el potencial de S. dasyphyllum como fuente genética para desarrollar variedades de berenjena tolerantes a la sequía, enfatizando la importancia de usar parientes silvestres para mejorar la tolerancia de los cultivos.
En general, esta tesis doctoral demuestra el potencial de los parientes silvestres de la berenjena para la mejora de la tolerancia a estreses abióticos. El desarrollo de retrocruzamientos avanzados a partir de diferentes parientes silvestres amplió la variación genética, mostrando efectos significativos de las dosis de nitrógeno en varios rasgos e identificando individuos transgresivos con características agronómicas mejoradas. La detección y validación de QTLs proporcionó conocimientos sobre la productividad de la berenjena, la eficiencia en el uso del nitrógeno y los mecanismos de respuesta a la sequía, contribuyendo a variedades de berenjena más sostenibles. / [CA] El canvi climàtic, impulsat per activitats humanes, intensifica l'estrès abiòtic en l'agricultura, afectant la productivitat i la salut de l'ecosistema. La creixent demanda de productes vegetals expandeix les terres agrícoles, empitjorant el canvi climàtic i la desertificació. L'estrès per sequera afecta greument la productivitat dels cultius, mentre que la fertilització excessiva amb nitrogen altera el cicle del nitrogen, causant problemes ambientals i de salut.
L'albergínia (Solanum melongena L.) és una hortalissa important, i l'ús de recursos genètics, inclosos els parents silvestres, és crucial per als programes que busquen millorar la tolerància a la sequera i l'eficiència en l'ús del nitrogen. Els avanços en seqüenciació i genotipat han millorat la nostra comprensió dels caràcters relacionats amb l'estrès en els cultius.
Aquesta tesi doctoral explora el potencial dels parents silvestres de l'albergínia per a millorar la tolerància als estressos abiòtics. L'objectiu és identificar noves variacions genètiques per a programes de millora, centrant-se en recursos genètics, regions genòmiques i gens candidats per a obtindre varietats d'albergínia més resilients mitjançant anàlisis genòmiques, transcriptòmiques i fenotípiques.
El capítol I avalua retrocreuaments avançats (ABs) d'albergínia i parents silvestres per a millorar l'eficiència en l'ús del nitrogen (NUE) en condicions de baix nitrogen (N). L'estudi de S. elaeagnifolium i els ABs en el fons genètic de l'albergínia en condicions de baix N va revelar diferències significatives entre els parentals. L'alta diversitat fenotípica en els ABs va mostrar alguns individus transgressius amb majors rendiments i NUE que el parental cultivat. Les avaluacions d'ABs de diferents bancs genètics (S. insanum, S. dasyphyllum i S. elaeagnifolium) en condicions de baix N van destacar el potencial d'aquestes espècies silvestres. L'alta diversitat en els ABs recolza el potencial dels parents silvestres per a millorar la resiliència de l'albergínia sota estrès abiòtic. Les dades de genotipat i fenotipat van permetre la identificació i validació de QTLs per a diversos caràcters, ampliant la comprensió genòmica de l'albergínia.
El capítol II investiga els mecanismes de resposta a la sequera en l'albergínia cultivada (S. melongena) i el seu parent silvestre S. dasyphyllum. Avaluats en condicions hidropòniques amb concentracions de polietilenglicol (PEG) en diferents etapes fenològiques, la seqüenciació d'ARN (RNA-Seq) va analitzar els patrons d'expressió gènica. S. dasyphyllum va demostrar una tolerància superior a l'estrès osmòtic en comparació amb S. melongena, amb més gens diferencialment expressats (DEGs) sota estrès. Les anàlisis d'enriquiment GO i les vies KEGG van revelar que ambdues espècies van activar diversos factors de transcripció i vies de resposta a l'estrès, amb S. dasyphyllum mostrant una regulació gènica més extensa. Les vies clau van incloure la senyalització d'ABA, la senyalització de MAPK i diverses vies de biosíntesi. Aquests resultats destaquen el potencial de S. dasyphyllum com a font genètica per a desenvolupar varietats d'albergínia tolerants a la sequera, emfatitzant la importància d'usar parents silvestres per a millorar la tolerància dels cultius.
En general, aquesta tesi doctoral demostra el potencial dels parents silvestres de l'albergínia per a la millora de la tolerància a estressos abiòtics. El desenvolupament de retrocreuaments avançats a partir de diferents parents silvestres va ampliar la variació genètica, mostrant efectes significatius de les dosis de nitrogen en diversos caràcters i identificant individus transgressius amb característiques agronòmiques millorades. La detecció i validació de QTLs va proporcionar coneixements sobre la productivitat de l'albergínia, l'eficiència en l'ús del nitrogen i els mecanismes de resposta a la sequera, contribuint a varietats d'albergínia més sostenibles. / [EN] Climate change, driven by human activities, intensifies abiotic stress in agriculture, impacting productivity and ecosystem health. Rising demands for plant products expand agricultural lands, worsening climate change and desertification. Drought stress severely affects crop productivity, while excessive nitrogen fertilization disrupts the nitrogen cycle, causing environmental and health issues.
Eggplant (Solanum melongena L.) is an important vegetable, and using genetic resources, including wild relatives, is crucial for breeding programs aimed at enhancing drought tolerance and nitrogen use efficiency. Advances in sequencing and genotyping have improved our understanding of stress-related traits in crops, supporting resilient cultivar development.
This doctoral thesis explores the potential of wild eggplant relatives to improve tolerance to abiotic stresses. The goal is to identify new genetic variations for breeding programs, focusing on valuable genetic resources, genomic regions, and candidate genes for more resilient eggplant varieties through genomic, transcriptomic, and phenotypic analyses.
Chapter I evaluates advanced backcrosses (ABs) of eggplant and wild relatives to improve nitrogen use efficiency (NUE) under low nitrogen (N) conditions. The study of S. elaeagnifolium and ABs in the eggplant genetic background under low N conditions revealed significant differences among parentals. High phenotypic diversity in ABs showed some transgressive individuals with higher yields and NUE than the cultivated parent. Evaluations of ABs from different genepools (S. insanum, S. dasyphyllum, and S. elaeagnifolium) under low N conditions highlighted the potential of these wild species for low nitrogen input breeding. Low N conditions decreased chlorophyll content but increased flavonol and anthocyanin levels, reducing aerial biomass, stem diameter, yield, and nitrogen and carbon content in plants and fruits. The high diversity in ABs supports the potential of wild relatives to enhance eggplant resilience under abiotic stress. Genotyping and phenotyping data enabled the identification and validation of QTLs for various traits, expanding the genomic understanding of eggplant.
Chapter II investigates drought response mechanisms in cultivated eggplant (S. melongena) and its wild relative S. dasyphyllum. Evaluated under hydroponic conditions with polyethylene glycol (PEG) concentrations (20% and 30%) at different phenological stages, RNA sequencing (RNA-Seq) analyzed gene expression patterns. S. dasyphyllum demonstrated superior tolerance to osmotic stress compared to S. melongena, with more differentially expressed genes (DEGs) under stress. GO enrichment and KEGG pathway analyses revealed that both species activated various transcription factors and stress response pathways, with S. dasyphyllum showing more extensive gene regulation. Key pathways included ABA signaling, MAPK signaling, and various biosynthesis pathways. These findings highlight S. dasyphyllum's potential as a gene source for developing drought-tolerant eggplant varieties, emphasizing the importance of using wild relatives to enhance crop tolerance and sustainability.
Overall, this doctoral thesis demonstrates the potential of wild eggplant relatives for breeding tolerance to abiotic stresses. Developing advanced backcrosses from different crop wild relatives expanded genetic variation, showing significant effects of nitrogen doses on various traits and identifying transgressive individuals with improved agronomic characteristics. The detection and validation of QTLs provided insights into eggplant productivity, nitrogen use efficiency, and drought response mechanisms, contributing to more resilient and sustainable eggplant varieties. / Villanueva Párraga, G. (2024). Evaluation of Eggplant, Wild Relatives and Introgression Breeding Materials for Tolerance to Abiotic Stresses [Tesis doctoral]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/209414 / Compendio
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