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Genomic selection (GS) has become an indispensable tool in modern plant breeding, particularly for complex traits. This study aimed to assess the efficacy of GS in predicting rust (Uromyces pisi) resistance in pea (Pisum sativum), using a panel of 320 pea accessions and a set of 26,045 Silico-Diversity Arrays Technology (Silico-DArT) markers. We compared the prediction abilities of different GS models and explored the impact of incorporating marker × environment (M×E) interaction as a covariate in the GBLUP (genomic best linear unbiased prediction) model. The analysis included phenotyping data from both field and controlled conditions. We assessed the predictive accuracies of different cross-validation strategies and compared the efficiency of using single traits versus a multi-trait index, based on factor analysis and ideotype-design (FAI-BLUP), which combines traits from controlled conditions. The GBLUP model, particularly when modified to include M×E interactions, consistently outperformed other models, demonstrating its suitability for traits affected by complex genotype-environment interactions (GEI). The best predictive ability (0.635) was achieved using the FAI-BLUP approach within the Bayesian Lasso (BL) model. The inclusion of M×E interactions significantly enhanced prediction accuracy across diverse environments in GBLUP models, although it did not markedly improve predictions for non-phenotyped lines. These findings underscore the variability of predictive abilities due to GEI and the effectiveness of multi-trait approaches in addressing complex traits. Overall, our study illustrates the potential of GS, especially when employing a multi-trait index like FAI-BLUP and accounting for M×E interactions, in pea breeding programs focused on rust resistance.

Abstract Background Rust is a damaging disease affecting vital crops, including pea, and identifying highly resistant genotypes remains a challenge. Accurate measurement of infection levels in large germplasm collections is crucial for finding new resistance sources. Current evaluation methods rely on visual estimation of disease severity and infection type under field or controlled conditions. While they identify some resistance sources, they are error-prone and time-consuming. An image analysis system proves useful, providing an easy-to-use and affordable way to quickly count and measure rust-induced pustules on pea samples. This study aimed to develop an automated image analysis pipeline for accurately calculating rust disease progression parameters under controlled conditions, ensuring reliable data collection. Results A highly efficient and automatic image-based method for assessing rust disease in pea leaves was developed using R. The method’s optimization and validation involved testing different segmentation indices and image resolutions on 600 pea leaflets with rust symptoms. The approach allows automatic estimation of parameters like pustule number, pustule size, leaf area, and percentage of pustule coverage. It reconstructs time series data for each leaf and integrates daily estimates into disease progression parameters, including latency period and area under the disease progression curve. Significant variation in disease responses was observed between genotypes using both visual ratings and image-based analysis. Among assessed segmentation indices, the Normalized Green Red Difference Index (NGRDI) proved fastest, analysing 600 leaflets at 60% resolution in 62 s with parallel processing. Lin’s concordance correlation coefficient between image-based and visual pustule counting showed over 0.98 accuracy at full resolution. While lower resolution slightly reduced accuracy, differences were statistically insignificant for most disease progression parameters, significantly reducing processing time and storage space. NGRDI was optimal at all time points, providing highly accurate estimations with minimal accumulated error. Conclusions A new image-based method for monitoring pea rust disease in detached leaves, using RGB spectral indices segmentation and pixel value thresholding, improves resolution and precision. It rapidly analyses hundreds of images with accuracy comparable to visual methods and higher than other image-based approaches. This method evaluates rust progression in pea, eliminating rater-induced errors from traditional methods. Implementing this approach to evaluate large germplasm collections will improve our understanding of plant-pathogen interactions and aid future breeding for novel pea cultivars with increased rust resistance.

Little resistance to the pea weevil insect pest (Bruchus pisorum) is available in pea (Pisum sativum) cultivars, highlighting the need to search for sources of resistance in Pisum germplasm and to decipher the genetic basis of resistance. To address this need, we screened the response to pea weevil in a Pisum germplasm collection (324 accession, previously genotyped) under field conditions over four environments. Significant variation for weevil seed infestation (SI) was identified, with resistance being frequent in P. fulvum, followed by P. sativum ssp. elatius, P. abyssinicum, and P. sativum ssp. humile. SI tended to be higher in accessions with lighter seed color. SI was also affected by environmental factors, being favored by high humidity during flowering and hampered by warm winter temperatures and high evapotranspiration during and after flowering. Merging the phenotypic and genotypic data allowed genome-wide association studies (GWAS) yielding 73 markers significantly associated with SI. Through the GWAS models, 23 candidate genes were found associated with weevil resistance, highlighting the interest of five genes located on chromosome 6. These included gene 127136761 encoding squalene epoxidase; gene 127091639 encoding a transcription factor MYB SRM1; gene 127097033 encoding a 60S ribosomal protein L14; gene 127092211, encoding a BolA-like family protein, which, interestingly, was located within QTL BpLD.I, earlier described as conferring resistance to weevil in pea; and gene 127096593 encoding a methyltransferase. These associated genes offer valuable potential for developing pea varieties resistant to Bruchus spp. and efficient utilization of genomic resources through marker-assisted selection (MAS).

Peas (Pisum sativum) are the fourth most cultivated pulses worldwide and a critical source of protein in animal feed and human food. Developing pea core collections improves our understanding of pea evolution and may ease the exploitation of their genetic diversity in breeding programs. We carefully selected a highly diverse pea core collection of 325 accessions and established their genetic diversity and population structure. DArTSeq genotyping provided 35,790 polymorphic DArTseq markers, of which 24,279 were SilicoDArT and 11,511 SNP markers. More than 90% of these markers mapped onto the pea reference genome, with an average of 2787 SilicoDArT and 1644 SNP markers per chromosome, and an average LD50 distance of 0.48 and 1.38 Mbp, respectively. The pea core collection clustered in three or six subpopulations depending on the pea subspecies. Many admixed accessions were also detected, confirming the frequent genetic exchange between populations. Our results support the classification of Pisum genus into two species, P. fulvum and P. sativum (including subsp. sativum, arvense, elatius, humile, jomardii and abyssinicum). In addition, the study showed that wild alleles were incorporated into the cultivated pea through the intermediate P. sativum subsp. jomardii and P. sativum subsp. arvense during pea domestication, which have important implications for breeding programs. The high genetic diversity found in the collection and the high marker coverage are also expected to improve trait discovery and the efficient implementation of advanced breeding approaches.

Pea rust is a major disease worldwide caused by Uromyces pisi in temperate climates. Only moderate levels of partial resistance against U. pisi have been identified so far in pea, urging for enlarging the levels of resistance available for breeding. Herein, we describe the responses to U. pisi of 320 Pisum spp. accessions, including cultivated pea and wild relatives, both under field and controlled conditions. Large variations for U. pisi infection response for most traits were observed between pea accessions under both field and controlled conditions, allowing the detection of genotypes with partial resistance. Simultaneous multi-trait indexes were applied to the datasets allowing the identification of partial resistance, particularly in accessions JI224, BGE004710, JI198, JI199, CGN10205, and CGN10206. Macroscopic observations were complemented with histological observations on the nine most resistant accessions and compared with three intermediates and three susceptible ones. This study confirmed that the reduced infection of resistant accessions was associated with smaller rust colonies due to a reduction in the number of haustoria and hyphal tips per colony. Additionally, a late acting hypersensitive response was identified for the first time in a pea accession (PI273209). These findings demonstrate that screening pea collections continues to be a necessary method in the search for complete resistance against U. pisi. In addition, the large phenotypic diversity contained in the studied collection will be useful for further association analysis and breeding perspectives.

In Mediterranean environments, with mild winters and dry summers, peas are planted in autumn or early winter to profit from winter rain and to avoid terminal drought and high summer temperatures. The root parasitic weed broomrape (Orobanche crenata) appears as a major limiting factor under these conditions. To address such specific growing conditions and associated constraints, targeted breeding is needed. We present here recent achievements in the development of pea lines arising from a wide hybridization program incorporating resistance to broomrape and to powdery mildew (Erysiphe pisi) from landraces and wild relatives. Their adaption to autumn sowings under Mediterranean rain fed conditions, and their agronomic performance and resistance to prevailing diseases is compared with those of check cultivars in a multi-environment field test with nine trials performed over three seasons. HA-GGE biplots were a powerful tool for comparison among accessions in terms of performance and stability for each trait assessed. Like this, breeding lines NS22, NS34, NS8, NS39, NS35, NS21 and NS83 over-yielded all check cultivars. Grain yield was strongly affected by broomrape infection, with little influence of powdery mildew and ascochyta blight. All breeding lines studied showed high to moderate resistance to broomrape, whereas all check cultivars were severely infected. Broomrape infection was not correlated with days to flowering, whereas powdery mildew infection was favored by long cycles. Broomrape infection was enhanced by mild winter temperatures before flowering and spring rain, whereas high spring temperatures hampered broomrape development.