Decreased stomatal density in poplar leads to improved water-use efficiency and drought tolerance

Regulation of stomatal density is a topic that has recently drawn a lot of attention worldwide. Stomata serve to control CO2 influx and water efflux from leaves and play a direct role in the global water cycle. This project researched genes that are able to increase or decrease leaf stomatal numbers when mutated or manipulated in a number of model species. The focus of this research was to generate highly drought tolerant trees with increased water conservation abilities in order to alleviate pressures placed on current water supplies. The goal of this project was to decrease stomatal numbers in poplar as a method of altering water usage and to achieve greater water-use efficiency. In Arabidopsis thaliana, epidermal patterning factor1 (EPF1) is involved in stomatal complex development and serves to control leaf stomatal number and spatial distribution. The most prominent role of EPF1 involves the enforcement of the one-cell-spacing rule that prevents multiple stomata from developing adjacent to each other without an epidermal or pavement cell for separation. Overexpression of AtEPF1 in Arabidopsis and poplar resulted in reduced stomatal density, a characteristic recently shown to be directly related to water-use efficiency and indirectly to drought tolerance. These experiments provided the background necessary to identify and isolate the poplar homolog of AtEPF1; PtaEPF1. PtaEPF1 was overexpressed in Arabidopsis and poplar and the resulting data indicated that both genes are able to decrease stomatal density and limited leaf water exodus from the leaves when overexpressed; however the transgenic plants generated in this study showed several abnormal leaf phenotypes in addition to decreased stomatal densities. Although no significant decreases in overall leaf biomass were detected in these plants, further decreases in stomatal density were likely to negatively influence CO2 assimilation rates. This study compiled currently available data regarding responses of plants to climate change, reforestation efforts, breeding technologies, and unconventional methods of generating abiotic tolerance to ensure this research was unique and provided valuable information to the research community. Leaf stomatal density data were collected from a selection of several fine hardwood tree species, Arabidopsis stomatal density mutants, and transgenics poplar generated during this analysis. This project began with a generalized idea regarding climate change and proceeded to narrow in focus from the analysis of numerous tree species to a single species outlook. Although often found in riparian areas, most Populus spp. are capable of tolerating a wide range of environments. Therefore, based on the data generated, the proposed improvements in abiotic tolerance would be most beneficial for a species unaccustomed to variable environmental conditions. Manipulation of stomatal density may allow endangered tree populations to tolerate the highly variable future climate events predicted to occur.

Insights into the Sulfate Transporter Gene Family and Its Expression Patterns in Durum Wheat Seedlings under Salinity

Sulfate transporters (SULTRs) are an essential plant transporter class responsible for the absorption and distribution of sulfur, an essential plant growth element. SULTRs are also involved in processes related to growth and development and in response to environmental stimuli. In the present study, 22 TdSULTR family members were identified and characterized in the genome of Triticum turgidum L. ssp. durum (Desf.) using available bioinformatics tools. The expression levels of candidate TdSULTR genes were investigated under salt treatments of 150 and 250 mM NaCl after several different exposure times. TdSULTRs showed diversity in terms of physiochemical properties, gene structure, and pocket sites. TdSULTRs and their orthologues were classified into the known five main plant groups of highly diverse subfamilies. In addition, it was noted that segmental duplication events could lengthen TdSULTR family members under evolutionary processes. Based on pocket site analysis, the amino acids leucine (L), valine (V), and serine (S) were most often detected in TdSULTR protein binding sites. Moreover, it was predicted that TdSULTRs have a high potential to be targeted by phosphorylation modifications. According to promoter site analysis, the plant bioregulators ABA and MeJA were predicted to affect TdSULTR expression patterns. Real-time PCR analysis revealed TdSULTR genes are differentially expressed at 150 mM NaCl but show similar expression in response to 250 mM NaCl. TdSULTR reached a maximum level of expression 72 h after the 250 mM salt treatment. Overall, we conclude that TdSULTR genes are involved in the response to salinity in durum wheat. However, additional studies of functionality are needed to determine their precise function and linked-interaction pathways

The First Complete Chloroplast Genome Sequence and Phylogenetic Analysis of Pistachio (Pistacia vera)

Pistachio is one of the most economically important nut crops worldwide. However, there are no reports describing the chloroplast genome of this important fruit tree. In this investigation, we assembled and characterized the complete pistachio chloroplast sequence. The Pistacia vera chloroplast genome was 160,598 bp in size, similar to other members of Anacardiaceae (149,011–172,199 bp) and exhibited the typical four section structure, including a large single copy region (88,174 bp), a small single copy region (19,330 bp), and a pair of inverted repeats regions (26,547 bp). The genome contains 121 genes comprised of 87 protein-coding genes, 30 tRNA genes and 4 rRNA genes. Thirteen intron-containing genes were identified in the genome wherein two genes had more than two introns. The genomic patterns of GC content resembled those for other Anacardiaceae. P. vera displayed the highest number of simple sequence repeats (SSRs) among the genera studied, which may be useful for molecular marker development and future population studies. Amino acid analysis revealed that Leucine is the most frequent (10.69%) amino acid in the chloroplast genome followed by Isoleucine (8.53%) and Serine (7.77%). Cysteine (1.30%) and Tryptophan (1.74%) were the least frequent amino acids. Phylogenetic analysis revealed P. vera is most like its taxonomically close relative P. weinmaniifolia, followed by Rhus chinensis; all placed taxonomically in the tribe Rhoeae. Members of Anacardiaceae were most closely related to Rhoeae, followed by members of Spondieae. The reports of this chloroplast genome will be useful for future conservation studies, genetic evaluation and breeding of P. vera, and more comprehensive phylogenetic analysis of the Pistacia species and its closely-related genera

Evaluation of genetic variability among “Early Mature” Juglans regia using microsatellite markers and morphological traits

Limiting the juvenile phase and reducing tree size are the two main challenges for breeders to improve most fruit crops. Early maturation and dwarf cultivars have been reported for many fruit species. “Early mature” and low vigor walnut genotypes were found among seedlings of Persian walnut. Nine microsatellite markers were used to evaluate genetic diversity among “Early Mature” Persian walnut accessions and provide a comparison with “normal growth” accessions. Six maturation related characteristics were also measured in “Early Mature” samples. Phenotypic traits and diversity indices showed relatively high levels of genetic diversity in “Early Mature” seedlings and indicated high differentiation between individuals. Seedling height, the most diverse phenotypic trait, has an important role in the clustering of “Early Mature” accessions. The “Early Mature” type had higher number of alleles, number of effective allele, and Shannon index compared to the “Normal Growth” group. The two types of studied walnuts had different alleles, with more than half of produced alleles specific to a specific group. “Early Mature” and “Normal Growth” walnuts had 27 and 17 private alleles, respectively. Grouping with different methods separated “Early Mature” and “Normal Growth” samples entirely. The presence of moderate to high genetic diversity in “Early Mature” walnuts and high genetic differentiation with “Normal Growth” walnuts, indicated that “Early Mature” walnuts were more diverse and distinct from “Normal Growth” samples. Moreover, our results showed SSR markers were useful for differentiating between “Early Mature” and “Normal Growth” walnuts. A number of identified loci have potential in breeding programs for identification of “Early Mature” walnuts at the germination phase

Genetic diversity and genetic structure of Persian walnut (Juglans regia) accessions from 14 European, African, and Asian countries using SSR markers

Persian walnut (Juglans regia L.) is the world’s most widely grown nut crop, but large-scale assessments and comparisons of the genetic diversity of the crop are notably lacking. To guide the conservation and utilization of Persian walnut genetic resources, genotypes (n = 189) from 25 different regions in 14 countries on three continents were sampled to investigate their genetic relationships and diversity using ten microsatellite (SSR) loci. The SSRs amplified from 3 to 25 alleles per locus, with a mean value of 11.5 alleles per locus. The mean values of observed and expected heterozygosity were 0.62 and 0.73, respectively. Based on Nei’s genetic identity, accessions from Bratislava (Slovakia) and Antalya (Turkey) showed the lowest similarity (0.36), while accessions from Algeria and Tunisia as well as accessions from Debrecen (Hungary) and Trnava (Slovakia) had the highest similarity (0.97). Two populations from Iran (Alborz and Ardabil) had the highest number of private alleles (7 and 5), but they were quite different as they also had the lowest genetic identity when compared to the remaining populations as well as to each other. Although overall differentiation among regions was relatively low (Fst = 0.07), cluster analysis grouped accessions generally but not completely according to geography. STRUCTURE software confirmed these results and divided the accessions into two main groups, separating accessions collected from Europe and North Africa from those from Greece and the Near East. Results indicate the presence of a likely center of diversity for Persian walnut in Eastern and Southeastern Europe. They also provide information that can be used to devise conservation actions. Notably, the genetic diversity of threatened populations from two regions in Iran should be conserved

Tracing Superior Late-Leafing Genotypes of Persian Walnut for Managing Late-Spring Frost in Walnut Orchards

Evaluating genetic diversity in walnut (Juglans regia L.) populations is a rapid approach used by walnut breeding programs to distinguish superior genotypes. The present study was conducted on the walnut population of Hamedan Province, one of the richest and most genetically diverse regions in Iran, during 2018–2019. After the initial screening, 47 genotypes were selected for further evaluation of pomological and phenological traits based on International Plant Genetic Resources Institute (IPGRI) descriptors. Nut and kernel weights among the selected genotypes ranged from 7.15 to 21.05 g and 3.0 to 10.8 g, respectively. Principal component analysis (PCA) categorized the genotypes into three distinct groups. Whereas the cluster analysis (CA) revealed the similarities and dissimilarities among the genotypes by identifying four major clusters. Spearman correlation analysis showed a positive correlation (p p < 0.01) between shell thickness (STH) and packing tissue thickness (PTT) with kernel percentage (KP) was observed. Lastly, 10 of 47 genotypes (TAL8, TAL9, TAL10, TAL14, TAL19, TAL22, TB2, TB4, TB6, and RDGH5) were considered superior. Superior genotypes were late-leafing (25–40 days after the standard) and displayed a lateral bearing (LB) habit with heavy nuts (12.52–16.82 g) and kernels (6.53–8.15 g), thin shells (1.06–1.25 mm), and lightly colored kernels

Tracing Superior Late-Leafing Genotypes of Persian Walnut for Managing Late-Spring Frost in Walnut Orchards

Evaluating genetic diversity in walnut (Juglans regia L.) populations is a rapid approach used by walnut breeding programs to distinguish superior genotypes. The present study was conducted on the walnut population of Hamedan Province, one of the richest and most genetically diverse regions in Iran, during 2018&ndash;2019. After the initial screening, 47 genotypes were selected for further evaluation of pomological and phenological traits based on International Plant Genetic Resources Institute (IPGRI) descriptors. Nut and kernel weights among the selected genotypes ranged from 7.15 to 21.05 g and 3.0 to 10.8 g, respectively. Principal component analysis (PCA) categorized the genotypes into three distinct groups. Whereas the cluster analysis (CA) revealed the similarities and dissimilarities among the genotypes by identifying four major clusters. Spearman correlation analysis showed a positive correlation (p &lt; 0.01) between nut weight (NWT), nut size, and kernel weight (KW), while a negative correlation (p &lt; 0.01) between shell thickness (STH) and packing tissue thickness (PTT) with kernel percentage (KP) was observed. Lastly, 10 of 47 genotypes (TAL8, TAL9, TAL10, TAL14, TAL19, TAL22, TB2, TB4, TB6, and RDGH5) were considered superior. Superior genotypes were late-leafing (25&ndash;40 days after the standard) and displayed a lateral bearing (LB) habit with heavy nuts (12.52&ndash;16.82 g) and kernels (6.53&ndash;8.15 g), thin shells (1.06&ndash;1.25 mm), and lightly colored kernels

Pollen flow and paternity in an isolated and non-isolated black walnut (Juglans nigra L.) timber seed orchard.

Artificial pollination of black walnut (Juglans nigra L.) is not practical and timber breeders have historically utilized only open-pollinated half-sib families. An alternate approach called "breeding without breeding," consists of genotyping open-pollinated progeny using DNA markers to identify paternal parents and then constructing full-sib families. In 2014, we used 12 SSR markers to genotype 884 open-pollinated half-sib progeny harvested from two clonal orchards containing 206 trees, comprised of 52 elite timber selections. Seed was harvested in 2011 from each of two ramets of 23 clones, one upwind and one downwind, based on prevailing wind direction from the west-southwest. One orchard was isolated from wild black walnut and composed of forward selections while the other orchard was adjacent to a natural forest containing mature black walnut composed of backward selections. Isolation significantly increased within-orchard pollination (85%) of the progeny from the isolated orchard compared to 42% from the non-isolated orchard. Neither prevailing wind direction nor seed tree position in the orchard affected paternity patterns or wild pollen contamination. Genetic diversity indices revealed that progeny from both orchards were in Hardy-Weinberg equilibrium with very little inbreeding and no selfing. A significant level of inbreeding was present among the forward selected parents, but not the first generation (backward selected) parents. Some orchard clones failed to sire any progeny while other clones pollinated upwards of 20% of progeny