Assessing heat tolerance in potatoes: Responses to stressful Texas field locations and controlled contrasting greenhouse conditions

In recent years, heat stress has affected potato production more frequently, resulting in lower marketable yields and reduced tuber quality. In order to develop heat-tolerant potatoes, it is necessary to select under heat-stress conditions and consider traits affected by heat stress. The Texas A&M Potato Breeding Program has selected potatoes under high-temperature stress for several decades. Ten potato cultivars, representing heat tolerant and sensitive clones based on past performance in Texas, were included in field trials for three years at the two main locations used by the Texas Breeding Program (Dalhart and Springlake, TX) to assess if the Texas field locations are suitable for heat tolerance screening. Both locations were confirmed as appropriate for heat stress screening. However, Springlake was a more stressful location since it had significantly lower yields of marketable tubers and increased percentages of tuber defects. Planting time did not have a significant effect at the most stressful location. The same ten potato clones were included in greenhouse experiments with contrasting temperatures (normal versus heat stress). There was confirmation that heat stress conditions resulted in significantly lower marketable yields, specific gravity, dormancy, and significantly higher percentages of tuber defects; however, significant differences existed between potato clones. Under heat stress conditions, Russet Burbank had a high percent of tubers with external defects, whereas Atlantic showed the highest percentage of internal defects (mainly internal heat necrosis). Vanguard Russet produced the highest marketable yield while maintaining a low percentage of external and internal defects. Russet Burbank and Atlantic were heat-sensitive controls for external and internal tuber defects, respectively. In contrast, Vanguard Russet can be used as a reliable heat-tolerant control. Including appropriate controls in heat stress studies will help identify clones with heat tolerance

Genomic regions associated with tuber traits in tetraploid potatoes and identification of superior clones for breeding purposes

In potato breeding, morphological tuber traits are important selection targets to meet the demands of the fresh and processing markets. Understanding the genetic basis of tuber traits should guide selection and improve breeding efficiencies. However, this is challenging in potato due to the complexity of the traits and the polyploid nature of the potato genome. High-throughput affordable molecular markers and new software specific for polyploid species have the potential to unlock previously unattainable levels of understanding of the genetic basis of tuber traits in tetraploid potato. In this study, we genotyped a diversity panel of 214 advanced clones with the 22 K SNP potato array and phenotyped it in three field environments in Texas. We conducted a genome-wide association study using the GWASpoly software package to identify genomic regions associated with tuber morphological traits. Some of the QTLs discovered confirmed prior studies, whereas others were discovered for the first time. The main QTL for tuber shape was detected on chromosome 10 and explained 5.8% of the phenotypic variance. GWAS analysis of eye depth detected a significant QTL on chromosome 10 and explained 3.9% of the phenotypic variance. Another QTL peak for eye depth on chromosome 5 was located near the CDF1 gene, an important regulator of maturity in potato. Our study found that multiple QTLs govern russeting in potato. A major QTL for flesh color on chromosome 3 that explained 26% of the phenotypic variance likely represents the Y locus responsible for yellow flesh in potato tubers. Several QTLs were detected for purple skin color on chromosome 11. Furthermore, genomic estimated breeding values were obtained, which will aid in the early identification of superior parental clones that should increase the chances of producing progenies with higher frequencies of the desired tuber traits. These findings will contribute to a better understanding of the genetic basis of morphological traits in potato, as well as to identifying parents with the best breeding values to improve selection efficiency in our potato breeding program

GGE biplot based assessment of yield stability, adaptability and mega-environment characterization for hybrid pigeon pea (Cajanus cajan

ABSTRACT GGE biplot methodology is a powerful tool to study relationship among test environments (E), genotypes (G) and genotype-by-environment interaction (GE). Present study was conducted on 10 short-duration genotypes in five test environments for two years, and 16 medium-duration genotypes in six test locations for three years in randomized complete block design with two replications. In short-maturity group three mega-environments (ME) were found-ME1 comprised of Phaltan, Patancheru and Hyderabad1; ME2 and 3 constituted Jalna and Aurangabad, respectively. In scenario of limited resources, Patancheru may be a good testing location for general adaptability of short-duration hybrids, while Aurangabad and Hyderabad1 may be right environments for testing specific adaptation of short-duration cultivars in pigeonpea. ICPH 2433 was a winning genotype in ME1 in terms of high yield and stability. In medium-maturity group, two MEs were observed. Jalna, Jalna 1, Parbhani and Hyderabad grouped together as ME1, while Patancheru and Phaltan formed the second mega-environment (ME2). Parbhani was found to be most representative of all the six test locations. Jalna (ME1) and Phaltan (ME2) produced longest environment vectors, and hence may be regarded as highly discriminating. In mediummaturity group ICPH 2673 was found to be stable and high-yielding genotype for ME1

CRISPR/Cas9-mediated glycolate oxidase disruption is an efficacious and safe treatment for primary hyperoxaluria type I

CRISPR/Cas9 technology offers novel approaches for the development of new therapies for many unmet clinical needs, including a significant number of inherited monogenic diseases. However, in vivo correction of disease-causing genes is still inefficient, especially for those diseases without selective advantage for corrected cells. We reasoned that substrate reduction therapies (SRT) targeting non-essential enzymes could provide an attractive alternative. Here we evaluate the therapeutic efficacy of an in vivo CRISPR/Cas9-mediated SRT to treat primary hyperoxaluria type I (PH1), a rare inborn dysfunction in glyoxylate metabolism that results in excessive hepatic oxalate production causing end-stage renal disease. A single systemic administration of an AAV8-CRISPR/Cas9 vector targeting glycolate oxidase, prevents oxalate overproduction and kidney damage, with no signs of toxicity in Agxt1(-/-) mice. Our results reveal that CRISPR/Cas9-mediated SRT represents a promising therapeutic option for PH1 that can be potentially applied to other metabolic diseases caused by the accumulation of toxic metabolites

CRISPR/Cas9-Mediated Mutagenesis of the Granule-Bound Starch Synthase Gene in the Potato Variety Yukon Gold to Obtain Amylose-Free Starch in Tubers

Potato (Solanum tuberosum L.) is the third most important food crop after rice and wheat. Its tubers are a rich source of dietary carbohydrates in the form of starch, which has many industrial applications. Starch is composed of two polysaccharides, amylose and amylopectin, and their ratios determine different properties and functionalities. Potato varieties with higher amylopectin have many food processing and industrial applications. Using Agrobacterium-mediated transformation, we delivered Clustered regularly interspaced short palindromic repeats and CRISPR-associated protein 9 (CRISPR/Cas9) reagents to potato (variety Yukon Gold) cells to disrupt the granule-bound starch synthase (gbssI) gene with the aim of eliminating the amylose component of starch. Lugol-Iodine staining of the tubers showed a reduction or complete elimination of amylose in some of the edited events. These results were further confirmed by the perchloric acid and enzymatic methods. One event (T2-7) showed mutations in all four gbss alleles and total elimination of amylose from the tubers. Viscosity profiles of the tuber starch from six different knockout events were determined using a Rapid Visco Analyzer (RVA), and the values reflected the amylopectin/amylose ratio. Follow-up studies will focus on eliminating the CRISPR components from the events and on evaluating the potential of clones with various amylose/amylopectin ratios for food processing and other industrial applications

Genetic Basis of Potato Tuber Defects and Identification of Heat-Tolerant Clones

Heat stress during the potato growing season reduces tuber marketable yield and quality. Tuber quality deterioration includes external (heat sprouts, chained tubers, knobs) and internal (vascular discoloration, hollow heart, internal heat necrosis) tuber defects, as well as a reduction in their specific gravity and increases in reducing sugars that result in suboptimal (darker) processed products (french fries and chips). Successfully cultivating potatoes under heat-stress conditions requires planting heat-tolerant varieties that can produce high yields of marketable tubers, few external and internal tuber defects, high specific gravity, and low reducing sugars (in the case of processing potatoes). Heat tolerance is a complex trait, and understanding its genetic basis will aid in developing heat-tolerant potato varieties. A panel of 217 diverse potato clones was evaluated for yield and quality attributes in Dalhart (2019 and 2020) and Springlake (2020 and 2021), Texas, and genotyped with the Infinium 22 K V3 Potato Array. A genome-wide association study was performed to identify genomic regions associated with heat-tolerance traits using the GWASpoly package. Quantitative trait loci were identified on chromosomes 1, 3, 4, 6, 8, and 11 for external defects and on chromosomes 1, 2, 3, 10, and 11 for internal defects. Yield-related quantitative trait loci were detected on chromosomes 1, 6, and 10 pertaining to the average tuber weight and tuber number per plant. Genomic-estimated breeding values were calculated using the StageWise package. Clones with low genomic-estimated breeding values for tuber defects were identified as donors of good traits to improve heat tolerance. The identified genomic regions associated with heat-tolerance attributes and the genomic-estimated breeding values will be helpful to develop new potato cultivars with enhanced heat tolerance in potatoes

Integrated physiological and molecular approaches to improvement of abiotic stress tolerance in two pulse crops of the semi-arid tropics

Chickpea (Cicer arietinum L.) and pigeonpea [Cajanus cajan L. (Millsp.)] play an important role in mitigating protein malnutrition for millions of poor vegetarians living in regions of the semi-arid tropics. Abiotic stresses such as excess and limited soil moisture (water-logging and drought), heat and chilling (high and low temperature stresses), soil salinity, and acidity are major yield constraints, as these two crops are grown mostly under rainfed conditions in risk-prone marginal and degraded lands with few or no inputs. Losses due to such stresses vary from 30% to 100% depending on their severity. The literature abounds in basic information concerning screening techniques, physiological mechanisms, and genetics of traits associated with resistance/tolerance to abiotic stresses in these two crops. However, the final outcome in terms of resistant/tolerant varieties has been far from satisfactory. This situation calls for improving selection efficiency through precise phenotyping and genotyping under high-throughput controlled conditions using modern tools of genomics. In this review, we suggest that an integrated approach combining advances from genetics, physiology, and biotechnology needs to be used for higher precision and efficiency of breeding programs aimed at improving abiotic stress tolerance in both chickpea and pigeonpea

Radiation Hybrid Mapping of the Species Cytoplasm-Specific (scs(ae)) Gene in Wheat

Radiation hybrid (RH) mapping is based on radiation-induced chromosome breakage and analysis of chromosome segment retention or loss using molecular markers. In durum wheat (Triticum turgidum L., AABB), an alloplasmic durum line [(lo) durum] has been identified with chromosome 1D of T. aestivum L. (AABBDD) carrying the species cytoplasm-specific (scs(ae)) gene. The chromosome 1D of this line segregates as a whole without recombination, precluding the use of conventional genome mapping. A radiation hybrid mapping population was developed from a hemizygous (lo) scs(ae)− line using 35 krad gamma rays. The analysis of 87 individuals of this population with 39 molecular markers mapped on chromosome 1D revealed 88 radiation-induced breaks in this chromosome. This number of chromosome 1D breaks is eight times higher than the number of previously identified breaks and should result in a 10-fold increase in mapping resolution compared to what was previously possible. The analysis of molecular marker retention in our radiation hybrid mapping panel allowed the localization of scs(ae) and 8 linked markers on the long arm of chromosome 1D. This constitutes the first report of using RH mapping to localize a gene in wheat and illustrates that this approach is feasible in a species with a large complex genome

Genetic architecture of tuber-bound free amino acids in potato and effect of growing environment on the amino acid content

Abstract Free amino acids in potato tubers contribute to their nutritional value and processing quality. Exploring the natural variation in their accumulation in tubers across diverse genetic backgrounds is critical to potato breeding programs aiming to enhance or partition their distribution effectively. This study assessed variation in the tuber-bound free amino acids in a diversity panel of tetraploid potato clones developed and maintained by the Texas A&M Potato Breeding Program to explore their genetic basis and to obtain genomic-estimated breeding values for applied breeding purposes. Free amino acids content was evaluated in tubers of 217 tetraploid potato clones collected from Dalhart, Texas in 2019 and 2020, and Springlake, Texas in 2020. Most tuber amino acids were not affected by growing location, except histidine and proline, which were significantly lower (− 59.0%) and higher (+ 129.0%), respectively, at Springlake, Texas (a location that regularly suffers from abiotic stresses, mainly high-temperature stress). Single nucleotide polymorphism markers were used for genome-wide association studies and genomic selection of clones based on amino acid content. Most amino acids showed significant variations among potato clones and moderate to high heritabilities. Principal component analysis separated fresh from processing potato market classes based on amino acids distribution patterns. Genome-wide association studies discovered 33 QTL associated with 13 free amino acids. Genomic-estimated breeding values were calculated and are recommended for practical potato breeding applications to select parents and advance clones with the desired free amino acid content