Climate change is one of the biggest challenges facing agriculture today. Transient or prolonged heat stress can be detrimental to plant reproductive development. The male gametophyte, pollen, is particularly sensitive to heat stress, resulting in sterile pollen (pre-anthesis) or deformed/stunted pollen tubes (post-anthesis). Quinoa (Chenopodium quinoa Willd.) has recently seen a rise in global interest due to its nutritional qualities, but global expansion of quinoa is partially hindered by its susceptibility to heat. It has been hypothesized that introgression of heat tolerance traits from wild relatives that occupy warmer environments can increase thermotolerance in quinoa. The goal of this research was to investigate the effect of heat stress on mature pollen grains from quinoa and its wild relatives, C. berlandieri and C. hircinum. To answer this question, several experiments were performed: 1. The nuclear number of the pollen of wild relatives was assessed to determine whether mature pollen grains were released at the trinucleate stage. All pollen was found to be trinucleate. 2. Pollen germination medium was optimized for four accessions: C. quinoa (QQ74; PI 614886), C. berlandieri var. zschackei (CB; BYU14118), and C. hircinum (CHA; Hircinum-069 and CHC; BYU17105). Optimal sucrose and PEG concentrations were determined to be: 5% sucrose/20% PEG for QQ74 and CB; 20% sucrose/0% PEG for CHA; and 10% sucrose/20% PEG for CHC. 3. Temperature optima for pollen germination for QQ74 was 32°C-36°C; CB was 30°C-34°C; CHA was 36°C; and CHC was 32°C -34°C. Overall, pollen from wild relatives was not found to be more heat-tolerant than pollen from domesticated quinoa. 4. Pollen tube elongation over time was observed for all four accessions at 34°C and 38°C, with CHA and QQ74 having the lowest decrease in rate at 38°C (35 and 45%, respectively). This study provides a new method for pollen collection for quinoa and its wild relatives, further optimizes the pollen germination media for QQ74, introduces pollen germination media for three wild accessions, and investigates the effect of heat stress on mature pollen grains. These observations can be employed in future studies investigating heat stress response of pollen in quinoa and its wild relatives.

We are now witnessing changing environmental conditions and these changes will likely continue into the coming decades due to projected increases in temperatures on the earth surface. Recent model projections suggest that the global mean surface air temperatures will increase by 1.4 to 5.8 °C by 2100. The consequences of these high temperatures include reductions in crop yields. Soybean is one of the major crops grown in the US, where high temperatures (>35 °C) during reproductive growth decreased yield. Two experiments were conducted to determine the effects of temperature on soybean pollen germination properties and identify responses to temperature among genotypes. Pollen collected from 44 genotypes grown outdoors in large pots was subjected to in vitro temperature treatments that varied from 15 to 50 °C at 5 °C increments in Experiment I. The mean cardinal temperatures (Tbase, Topt, and Tmax), averaged over 44 genotypes, were 13.2, 30.2 and 47.2 °C for pollen germination and 12.1, 36.1 and 47.0 °C for pollen tube growth. The relationship between Tbase and Topt for pollen germination was high indicating that genotypes with high Tbase also had higher Topt. The relationship was weak between Tbase and Tmax. The Topt for pollen tube growth was 5.8 ¢XC higher than the Topt for pollen germination. The study also showed significant genotypic variability for physiological parameters studied, but no significant correlations between the observed physiological parameters with any of the pollen germination or pollen tube growth parameters were studied. This indicated that physiological parameters, measured under ambient conditions may not be useful to identify reproductive tolerance to high temperatures in soybean. Total response index (TSRI), the sum of individual responses of all pollen parameters differentiated genotypes and their tolerance to high temperature. A heat-tolerant genotype (DG 5630RR) identified using TSRI technique, when grown at optimum and high temperature conditions in experiment II, were actually less sensitive to high temperatures compared to heat-sensitive and heat-intermediate sensitive genotypes indicating that pollen can be used as screening tool for heat tolerance. The identified high-temperature tolerant genotypes, based on pollen germination parameters, might be useful in current and future soybean breeding programs.

Thesis (Ph.D.) -- Mississippi State University. Department of Plant and Soil Sciences. / Title from title screen. Includes bibliographical references.

The period between pollination and fertilization, or the progamic phase, is a critical life history stage in seed plants and innovations in this life history stage are hypothesized to have played an important role in the diversification of flowering plants. Over the course of this dissertation research, I investigated programic phase development in Nymphaeales (water lilies), an ancient angiosperm lineage that diverged from the basalmost or next most basal node of the angiosperm phylogenetic tree and that is represented in the oldest angiosperm fossil record. I used field experiments and microscopy to document pollination biology, breeding system, and reproductive developmental traits in two families of Nymphaeales: Cabombaceae (Brasenia, Cabomba) and Hydatellaceae (Trithuria). Nymphaeales exhibits considerable variation in reproductive traits and true carpel closure, wind-pollination, and a primarily selfing breeding system have arisen independently in the lineage. Pollen tube pathway length, timing of stigma receptivity, and pollen tube growth rates are conspicuous traits that have undergone considerable modification in concert with shifts in pollination biology and breeding system. Post-pollination developmental processes in Nymphaeales appear to experience selective pressures similar to those experienced by more derived angiosperms and to evolve in similar ways. Nymphaeales also exhibits traits, such as accelerated pollen tube growth, callosic pollen tube walls, and the formation of callose plugs, that are almost certainly plesiomorphic in angiosperms and may have facilitated modification of carpel structure and progamic phase ontogenies. The finding that pollen tube traits that underlie developmental flexibility were already in place before the divergence of Nymphaeales supports the hypothesis that innovations in male gametophyte development were instrumental in facilitating early angiosperm diversification.

Kyoto University (京都大学) / 0048 / 新制・課程博士 / 博士(農学) / 甲第19043号 / 農博第2121号 / 新制||農||1032 / 31994 / 京都大学大学院農学研究科地域環境科学専攻 / (主査)教授 縄田 栄治, 教授 田中 千尋, 教授 北島 宣 / 学位規則第4条第1項該当

Chez les eucaryotes, les RNA Binding Proteins (RBP) s’associent aux transcrits pour former des Particules Ribo-Nucléoprotéiques (mRNP) dynamiques, dont la localisation et la composition sont déterminantes pour la maturation, l’export, la stabilité et la traduction des ARNm. Les protéines à motif LA sont des protéines de liaison à l’ARN, présentes chez plusieurs centaines d’espèces eucaryotes, qui se répartissent en 5 sous-familles : LA authentiques, LARP1, 4, 6 et 7. Les membres de ces sous-familles partagent des caractéristiques évolutives, des domaines additionnels et des fonctions conservés. Mes travaux de thèse ont contribué à l’étude fonctionnelle de protéines LARP6 chez Arabidopsis thaliana. On sait, à l’heure actuelle, que chez Manduca sexta et plusieurs espèces de vertébrés, LARP6 est impliquée dans la régulation de la différenciation cellulaire. Chez l’Homme, elle agit en tant que RBP pour coordonner la traduction des ARNm codant les sous-unités du collagène de type I. Les plantes vasculaires ont la particularité de contenir plusieurs protéines LARP6, classées en trois groupes évolutifs. Chez A. thaliana, l’unique représentant de chaque sous-famille semble s’être spécialisé. D’ailleurs, les protéines LARP6b et c ont des profils d’expression mutuellement exclusifs, où LARP6c est présente dans le pollen et LARP6b est ubiquitaire mais absente du gamétophyte mâle. Nous avons axé notre travail sur la protéine LARP6c et démontré qu’elle est cytoplasmique et impliquée, dans le pollen, dans le contrôle de la quantité d’ARNm codant des acteurs du transport vésiculaire. Les ARNm identifiés comme cibles potentielles de LARP6c codent, eux aussi, des facteurs impliqués dans le transport ; et possèdent dans leur 5’-UTR deux motifs qui pourraient permettre leur co-régulation par fixation de RBP. La délétion de LARP6c, affecte la capacité du tube pollinique à se diriger vers l’ovule suggérant un défaut de communication; ce qui est cohérent avec la dérégulation des ARNm codant des acteurs de la sécrétion/réception de signaux extracellulaires. Nous proposons que LARP6c intervient, dans le pollen, en tant que protéine de mRNP et co-régule la traduction et/ou la stabilité de transcrits codant des acteurs des voies de communications dépendantes de la sécrétion et de l’endocytose, et intervenant dans les échanges mâle/femelle / In eucaryotes, RNA Binding Proteins (RBP) associate with transcripts to form dynamic Ribo-Nucleoprotein Particles (mRNP), whose localization and composition are determinant for mRNA maturation, export, stability and translation. LA motif proteins are RNA binding proteins, found in several hundred eucaryotic species, which fall in 5 sub-families: genuine LA, LARP1, 4, 6 and 7. Members of these subfamilies share conserved evolutionary history, additional motifs and functions. My thesis work contributed to deciphering the functional properties of the Arabidopsis thaliana LARP6 proteins. Currently, we know that in Manduca sexta and many vertebrates species LARP6 is implicated in the regulation of cellular differentiation. In humans, it acts as an RBP to coordinate the translation of mRNA coding for type I collagen subunits. Vascular plants differ in possessing many LARP6 proteins classified in three evolutionary groups. In A. thaliana, the unique member of each subfamily seems to be specialized. LARP6b and c proteins present mutually exclusive expression profiles, with LARP6c only present in pollen and LARP6b ubiquitously expressed except in the male gametophyte. We mostly focused our work on LARP6c and showed it to be cytoplasmic and implicated in controlling the level of mRNAs encoding vesicular transport actors in pollen tubes. Putative identified LARP6c mRNA baits also encode proteins involved in transport and share two motifs in their 5’-UTR that could allow their co-regulation via RBP binding. LARP6c deletion induces deficiencies in pollen tube guidance towards the ovule, suggesting a communication default. This is consistent with the deregulation of mRNA coding for extra-cellular signal secretion/reception actors. We propose that LARP6c acts as an mRNP protein in pollen and co-regulates translation and/or stability of mRNA coding for actors of communication pathways depending on secretion and endocytosis; hence acting on male/female exchanges.

Nitric oxide (NO) is an important signaling molecule in plants. In the pollen of Arabidopsis thaliana, NO causes re-orientation of the growing tube and this response is mediated by 3′,5′-cyclic guanosine monophosphate (cGMP). However, in plants, NO-sensors have remained somewhat elusive. Here, the findings of an NO-binding candidate, Arabidopsis thaliana DIACYLGLYCEROL KINASE 4 (ATDGK4; AT5G57690) is presented. In addition to the annotated diacylglycerol kinase domain, this molecule also harbors a predicted heme-NO/oxygen (H-NOX) binding site and a guanylyl cyclase (GC) catalytic domain which have been identified based on the alignment of functionally conserved amino acid residues across species. A 3D model of the molecule was constructed, and from which the locations of the kinase catalytic center, the ATP-binding site, the GC and H-NOX domains were estimated. Docking of ATP to the kinase catalytic center was also modeled. The recombinant ATDGK4 demonstrated kinase activity in vitro, catalyzing the ATP-dependent conversion of sn-1,2-diacylglycerol (DAG) to phosphatidic acid (PA). This activity was inhibited by the mammalian DAG kinase inhibitor R59949 and importantly also by the NO donors diethylamine NONOate (DEA NONOate) and sodium nitroprusside (SNP). Recombinant ATDGK4 also has GC activity in vitro, catalyzing the conversion of guanosine-5'-triphosphate (GTP) to cGMP. The catalytic domains of ATDGK4 kinase and GC may be independently regulated since the kinase but not the GC, was inhibited by NO while Ca2+ only stimulates the GC. It is likely that the DAG kinase product, PA, causes the release of Ca2+ from the intracellular stores and Ca2+ in turn activates the GC domain of ATDGK4 through a feedback mechanism. Analysis of publicly available microarray data has revealed that ATDGK4 is highly expressed in the pollen. Here, the pollen tubes of mis-expressing atdgk4 recorded slower growth rates than the wild-type (Col-0) and importantly, they showed altered NO responses. Specifically, the mis-expressing atdgk4 pollen tubes have growth rates that were less affected by NO and showed reduced bending angles when challenged by an NO source. Further works on atdgk4 knockout/knockdown mutants will reveal the biological functions of ATDGK4 in NO and/or cGMP signaling in the pollen, and in the broader fertilization process.

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第19043号 / 農博第2121号 / 新制||農||1032(附属図書館) / 学位論文||H27||N4925(農学部図書室) / 31994 / 京都大学大学院農学研究科地域環境科学専攻 / (主査)教授 縄田 栄治, 教授 田中 千尋, 教授 北島 宣 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

No description available.

Invasive plants can alter pollination dynamics in invaded communities by disrupting patterns of pollinator visitation, pollen transfer dynamics (conspecific [CP] and heterospecific [HP]), and reproductive success. The direction of invasive effects (competitive, neutral, and facilitative) may be partially determined by spatial scale and species’ floral traits. Here, we investigated pollinator visitation, CP and HP receipt, and pollen tube growth for species in a C. arvense present community and non-present community at two scales. At the community-level, the effect of C. arvense on pollinator visitation varied among species. Floral symmetry seemed to explain this variation. At the floral neighborhood-level, we found competitive effects for pollinator visits and mixed effects on CP deposition. The overall structure of plant-plant HP deposition networks was slightly altered. We observed lower average centrality across shared species in the C. arvense present community suggesting C. arvense had subverted their roles as pollen donors.