Genome-wide association study for in vitro digestibility and related traits in triticale forage.

BACKGROUND: Triticale is making its way on dairy farms as an alternative forage crop. This requires the availability of high-yielding triticale varieties with good digestibility. Triticale forage breeding mainly focussed on biomass yield, but efforts to improve digestibility are increasing. We previously investigated the interrelationships among different quality traits in soft dough triticale: starch, acid detergent fibre and in vitro digestibility of organic matter (IVOMD) and of neutral detergent fibre (IVNDFD) of the total plant, IVNDFD and Klason lignin of the stems, and ear proportion and stem length. Here we determine the genetic control of these traits, using a genome-wide association (GWAS) approach. A total of 33,231 DArTseq SNP markers assessed in a collection of 118 winter triticale genotypes, including 101 varieties and 17 breeding lines, were used. RESULTS: The GWAS identified a total of 53 significant marker-trait associations (MTAs). The highest number of significantly associated SNP markers (n = 10) was identified for total plant IVNDFD. A SNP marker on chromosome 1A (4211801_19_C/T; 474,437,796 bp) was found to be significantly associated with ear proportion, and plant and stem IVNDFD, with the largest phenotypic variation for ear proportion (R²p = 0.23). Based on MTAs, candidate genes were identified which were of particular relevance for variation in in vitro digestibility (IVD) because they are putatively involved in plasma membrane transport, cytoskeleton organisation, carbohydrate metabolic processes, protein phosphorylation, and sterol and cell wall biogenesis. Interestingly, a xyloglucan-related candidate gene on chromosome 2R, SECCE2Rv1G0126340, was located in close proximity of a SNP significantly associated with stem IVNDFD. Furthermore, quantitative trait loci previously reported in wheat co-localized with significantly associated SNP markers in triticale. CONCLUSIONS: A collection of 118 winter triticale genotypes combined with DArTseq SNP markers served as a source for identifying 53 MTAs and several candidate genes for forage IVD and related traits through a GWAS approach. Taken together, the results of this study demonstrate that the genetic diversity available in this collection can be further exploited for research and breeding purposes to improve the IVD of triticale forage.

An insight into tissue culture-induced variation origin shared between anther culture-derived triticale regenerants.

BACKGROUND: The development of the plant in vitro techniques has brought about the variation identified in regenerants known as somaclonal or tissue culture-induced variation (TCIV). S-adenosyl-L-methionine (SAM), glutathione (GSH), low methylated pectins (LMP), and Cu(II) ions may be implicated in green plant regeneration efficiency (GPRE) and TCIV, according to studies in barley (Hordeum vulgare L.) and partially in triticale (× Triticosecale spp. Wittmack ex A. Camus 1927). Using structural equation models (SEM), these metabolites have been connected to the metabolic pathways (Krebs and Yang cycles, glycolysis, transsulfuration), but not for triticale. Using metabolomic and (epi)genetic data, the study sought to develop a triticale regeneration efficiency statistical model. The culture's induction medium was supplemented with various quantities of Cu(II) and Ag(I) ions for regeneration. The period of plant regeneration has also changed. The donor plant, anther-derived regenerants, and metAFLP were utilized to analyze TCIV concerning DNA in symmetric (CG, CHG) and asymmetric (CHH) sequence contexts. Attenuated Total Reflectance-Fourier Transfer Infrared (ATR-FTIR) spectroscopy was used to gather the metabolomic information on LMP, SAM, and GSH. To frame the data, a structural equation model was employed. RESULTS: According to metAFLP analysis, the average sequence change in the CHH context was 8.65%, and 0.58% was de novo methylation. Absorbances of FTIR spectra in regions specific for LMP, SAM, and GSH were used as variables values introduced to the SEM model. The average number of green regenerants per 100 plated anthers was 2.55. CONCLUSIONS: The amounts of pectin demethylation, SAM, de novo methylation, and GSH are connected in the model to explain GPRE. By altering the concentration of Cu(II) ions in the medium, which influences the amount of pectin, triticale's GPRE can be increased.

CRISPR/Cas9-mediated мultiplexed multi-allelic mutagenesis of genes located on A, B and R subgenomes of hexaploid triticale.

KEY MESSAGE: The first-time generation of hexaploid triticale plants harbouring variable panels of novel mutations in gene families involved in starch biosynthesis has been achieved by the subgenome-independent multiplexed CRISPR/Cas9-mediated editing.

Replacing the forage portion of the ration with triticale hay improves the performance of Holstein dairy cows.

The aim of the present study was to assess the effect of replacing the forage portion (alfalfa, corn silage, and barley straw) in the diet of lactating Holstein cows with triticale hay (TH, × Triticosecale L.) on DMI, digestibility, ruminal fermentation variables, estimated microbial-N synthesis (EMNS), and milk production and composition. Eight Holstein cows were used in a replicated Latin square design (two 4 × 4 squares) with four 28-d periods and 4 treatments, including a TH-free diet (control), and diets replacing 33%, 66%, and 100% of the forage portion with TH. Cows were fed ad libitum with 10% carryover during the experimental periods. Intakes and in vivo digestibilities of dry matter, organic matter, crude protein, and ash-free neutral detergent fiber, rumen pH, ammonia-N, total and individual short-chain fatty acids, protozoa, and bacteria populations were evaluated using specific methods. Moreover, in vitro total gas and methane release and in vivo urinary purine derivatives, EMNS, milk production, and composition were measured. The results showed that TH diets lowered DMI, compared with the control. Apparent digestibilities of DM, OM, CP, and NDF increased with dietary TH inclusion. The addition of TH instead of the diet forage portion increased in vivo rumen pH; acetic, propionic, valeric, and isovaleric acids concentrations; cellulolytic bacteria number; and in vitro gas production. In vivo rumen ammonia-N, short-chain fatty acids, butyric acid, in vivo and in vitro total protozoa and Entodiniinae numbers, and in vitro methane production decreased with increasing dietary levels of TH instead of the forage portion. The dietary addition of TH did not affect milk yield, protein, and lactose, but increased fat-corrected milk, milk fat, fat-corrected milk:DMI ratio, and milk yield:DMI ratio. Milk urea N decreased, but urinary purine derivatives excretion and EMNS increased with increasing levels of TH in the diet. For variables with significant changes, except for isovaleric acid, there was a linear response of animals to increasing levels of TH in the diet. Results suggest that TH, which is grown with less water compared with alfalfa, corn forage, and straw, is a potential alternative to those forages by increasing milk production efficiency, milk fat, and decreasing methane emission.

Temperature-Caused Changes in Raman Pattern and Protein Profiles of Winter Triticale (x Triticosecale, Wittm.) Field-Grown Seedlings.

Climate change, which causes periods with relatively high temperatures in winter in Poland, can lead to a shortening or interruption of the cold hardening of crops. Previous research indicates that cold acclimation is of key importance in the process of acquiring cereal tolerance to stress factors. The objective of this work was to verify the hypothesis that both natural temperature fluctuations and the plant genotype influence the content of metabolites as well as proteins, including antioxidant enzymes and photosystem proteins. The research material involved four winter triticale genotypes, differing in their tolerance to stress under controlled conditions. The values of chlorophyll a fluorescence parameters and antioxidant activity were measured in their seedlings. Subsequently, the contribution of selected proteins was verified using specific antibodies. In parallel, the profiling of the contents of chlorophylls, carotenoids, phenolic compounds, and proteins was carried out by Raman spectroscopy. The obtained results indicate that a better PSII performance along with a higher photosystem II proteins content and thioredoxin reductase abundance were accompanied by a higher antioxidant activity in the field-grown triticale seedlings. The Raman studies showed that the cold hardening led to a variation in photosynthetic dyes and an increase in the phenolic to carotenoids ratio in all DH lines.

Productive performance, digestibility, carcass traits and meat quality in rabbits fed triticale-based diets supplemented with xylanase.

Cereals such as triticale may contain high levels of xylans and arabinoxylans, limiting its use in diets since they act as anti-nutritional factors. The objective was to evaluate the effects of the enzyme xylanase included in triticale-based diets on productive performance, digestibility, carcass traits and meat quality in growing-finishing rabbits. Eighty rabbits (New Zealand X California breed), 35 days old, with an average initial live weight of 821 ± 26 g, were used. Twenty animals for treatment were used in each one of the fourth experimental treatments: 0, 4000, 8000 and 12,000 XU/kg of xylanase inclusion (XilaBlend 6X). The rabbits were fed ad libitum and fecal excretion was collected on days 7, 14, 21, 28 and 35 of the experimental period. At the end of the experimental period, the rabbits were slaughtered and carcass characteristics and meat quality were measured. A higher (P < 0.05) live weight was observed in rabbits fed diets with the addition of xylanase enzyme on days 4 and 7 of the experimental period. On the other hand, in the average total tract digestibility of organic matter, no significant difference was observed, similar to what occurred in the carcass traits and nutritional quality of the meat. The inclusion of 8000 XU/kg of xylanase enzyme provided the best values of apparent digestibility of total tract protein and dry matter on the finished stage of rabbits.

Investigation of chromosomal genetic characteristics and identification of structural variation in the offspring of hexaploid triticale×hexaploid wheat.

Hexaploid triticale is an important genetic resource for genetic improvement of common wheat, which can broaden the genetic basis of wheat. In order to lay a foundation for the subsequent research and utilization of triticale germplasm materials, the chromosomal genetic characteristics of cross and backcross offspring of hexaploid triticale×hexaploid wheat were investigated in the process of transferring rye chromatin from hexaploid triticale to hexaploid wheat. Hybrid and backcross combinations were prepared with hexaploid triticale 16yin171 as the maternal parent and hexaploid wheat Chuanmai62 as the paternal parent. The chromosomes in root tip cells of F1, BC1F1 and BC1F2 plants were traced and identified non-denaturing florescence in situ hybridization (ND-FISH). The results indicated that the backcross setting rate of hybrid F1 was 2.61%. The transmission frequency of 2R chromosome was the highest in BC1F1 plants while the transmissibility of rye chromosome in BC1F2 plant was 6R>4R>2R, and the 5B-7B wheat translocation in BC1F2 plants showed severe segregation. A total of 24 structural variant chromosomes were observed both in BC1F1 and BC1F2 plants, including chromosome fragments, isochromosomes, translocations, and dicentric chromosomes. In addition, the seed length and 1000-grain weight of some BC1F2 plants were better than that of the hexaploid wheat parent Chuanmai 62. Therefore, multiple backcrosses should be adopted as far as possible to make the rapid recovery of group D chromosomes, ensuring the recovery of fertility in offspring, when hexaploid tritriale is used as a bridge to introduce rye genetic material into common wheat. At the same time, the potential application value of chromosomal structural variation materials should be also concerned.

Triticale field phenotyping using RGB camera for ear counting and yield estimation.

Triticale (X Triticosecale Wittmack), a wheat-rye small grain crop hybrid, combines wheat and rye attributes in one hexaploid genome. It is characterized by high adaptability to adverse environmental conditions: drought, soil acidity, salinity and heavy metal ions, poorer soil quality, and waterlogging. So that its cultivation is prospective in a changing climate. Here, we describe RGB on-ground phenotyping of field-grown eighteen triticale market-available cultivars, made in naturally changing light conditions, in two consecutive winter cereals growing seasons: 2018-2019 and 2019-2020. The number of ears was counted on top-down images with an accuracy of 95% and mean average precision (mAP) of 0.71 using advanced object detection algorithm YOLOv4, with ensemble modeling of field imaging captured in two different illumination conditions. A correlation between the number of ears and yield was achieved at the statistical importance of 0.16 for data from 2019. Results are discussed from the perspective of modern breeding and phenotyping bottleneck.

Influence of Cuticular Waxes from Triticale on Rumen Fermentation: A Metabolomic and Microbiome Profiling Study.

Cuticular wax, a critical defense layer for plants, remains a relatively unexplored factor in rumen fermentation. We investigated the impact of cuticular wax on rumen fermentation using triticale as a model. In total, six wax classes were identified, including fatty acids, aldehydes, alkane, primary alcohol, alkyresorcinol, and ß-diketone, with low-bloom lines predominated by 46.05% of primary alcohols and high-bloom lines by 35.64% of ß-diketone. Low-wax addition (2.5 g/kg DM) increased the gas production by 19.25% (P < 0.05) and total volatile fatty acids by 6.34% (P > 0.05), and enriched key carbohydrate-fermenting rumen microbes like Saccharofermentans, Ruminococcus, and Prevotellaceae, when compared to non-wax groups. Metabolites linked to nucleotide metabolism, purine metabolism, and protein/fat digestion in the rumen showed a positive correlation with low-wax, benefiting rumen microbes. This study highlights the intricate interplay among cuticular wax, rumen microbiota, fermentation, and metabolomics in forage digestion, providing insights into livestock nutrition and forage utilization.

Response of nine triticale genotypes to different salt concentrations at the germination and early seedling stages.

Salinity stress poses a major challenge to agricultural productivity worldwide, and understanding their responses at the early growth stage is vital for devising strategies to cope with this stress. Therefore, to improve triticale productivity, this study investigated the salinity stress tolerance of different salt-tolerant triticale genotypes aiming to cultivate them on saline soils. To this end, salinity stress impacts on nine triticale genotypes, i.e., Zhongsi 1084, Gannong No. 2, Gannong No. 4, Shida No. 1, C6, C16, C23, C25 and C36 at germination and early seedling stages was evaluated. Each genotype was subjected to six treatments inducing control, 40, 80, 120, 160, and 200 mM NaCl treatments to study their effect on seedling and termination traits of the nine genotypes. Compared to the overall mean seedling vigor index, the seedling vigor index was higher in the genotypes Zhongsi 1084 and C6 (39% and 18.1%, respectively) and lower in Gannong No.2 (41%). Increasing NaCl concentrations negatively affected germination and seedling traits. Compared to other genotypes, Zhongsi 1084 had the highest mean germination rate, germination vigor index, germination percentage, mean daily germination and germination energy. It also showed the lowest relative salt injury. The relative salt injury was higher in the genotype Shida No. 1 than those in Gannong No. 2, Gannong No. 4, Shida No. 1, C16, and C36 genotypes. All genotypes exhibited desirable mean germination time except for line C6. High significant positive correlations were observed among germination rate, germination vigor index, germination percentage, mean daily germination, seedling vigor index, and root length. Principal component analysis (PCA) grouped the most desirable genotypes into two clusters. Our study determined salt stress tolerance of nine triticale genotypes at germination and early seedling stages. to select salt-tolerant genotypes that can be cultivated on saline soil or after salt irrigation.

Determining the proper sowing time for the mixture of Hungarian vetch and triticale under continental climate conditions / Determinação da época de semeadura adequada para a mistura de ervilhaca Húngara e triticale em condições climáticas continentais

The research was conducted to determine forage yield and some quality characteristics of Hungarian vetch + triticale mixture, sowed in five different times under rainfed conditions of Central Anatolia, Turkey. The mixture was sowed in the second, third and fourth week of October, and the first and the second week of November in 2017 and 2018. Depending on the sowing times, plant height (PH) of Hungarian vetch and triticale was between 46.7 and 59.4 cm, and 85.9 and 93.4 cm, respectively. Green forage yield (GFY) was between 1746.2 and 2059.4 kg da-1, dry matter yield (DMY) was between 541.0 and 707.6 kg da-1, crude protein yield (CPY) was between 80.4 and 110.3 kg da-1, digestible dry matter yield (DDMY) was between 340.8 and 453.9 kg da-1, acid detergent fiber (ADF) ratio was between 31.8 and 33.7%, neutral detergent fiber (NDF) ratio was between 44.7 and 49.5%, total digestible nutrient (TDN) was between 57.9 and 60.4% and relative feed value (RFV) was between 118.6 and 133.8. Sowing time had a significant effect (P < 0.05) on PH of triticale, while it has a very significant effect (P < 0.01) on GFY, DMY, CPY, DDMY, NDF ratios and RFV. Delaying the sowing time caused a decrease in the GFY, DMY and quality of the mixture. Results revealed that the first week of October is the most appropriate sowing time to obtain high dry matter yield with high quality under continental climate conditions of the Central Anatolia.

A pesquisa foi conduzida para determinar a produção de forragem e algumas características de qualidade da mistura de ervilhaca húngara + triticale, semeada em cinco épocas diferentes em regime de sequeiro na região Central da Anatólia, Turquia. A mistura foi semeada na segunda, terceira e quarta semanas de outubro, e na primeira e segunda semanas de novembro de 2017 e 2018. Dependendo da época de semeadura, a altura da planta (PH) da ervilhaca e triticale húngara foi entre 46,7 e 59,4 cm e 85,9 e 93,4 cm, respectivamente. O rendimento de forragem verde (GFY) ficou entre 1746,2 e 2059,4 kg da-1, o rendimento de matéria seca (DMY) foi entre 541,0 e 707,6 kg da-1, o rendimento de proteína bruta (CPY) foi entre 80,4 e 110,3 kg da-1, digestível seco o rendimento de matéria (DDMY) estava entre 340,8 e 453,9 kg da-1, a proporção da fibra em detergente ácido (FDA) estava entre 31,8 e 33,7%, a proporção da fibra em detergente neutro (FDN) estava entre 44,7 e 49,5%, o nutriente digestível total (NDT) era entre 57,9 e 60,4% e o valor relativo da alimentação (RFV) foi entre 118,6 e 133,8. A época de semeadura teve um efeito significativo (P < 0,05) no PH do triticale, enquanto teve um efeito muito significativo (P < 0,01) nas relações GFY, DMY, CPY, DDMY, FDN e RFV. O atraso na semeadura ocasionou diminuição no GFY, DMY e na qualidade da mistura. Os resultados revelaram que a primeira semana de outubro é a época de semeadura mais adequada para a obtenção de alta produtividade de matéria seca com alta qualidade nas condições climáticas continentais da Anatólia Central.

Triticale silage replaces sorghum silage in finishing of Braford heifers / Silagem de triticale substitui silagem de sorgo na terminação de novilhas da raça Braford

The aim of this study was to evaluate the effect of sorghum silage substitution with different proportions of triticale silage on the performance and carcass of Braford heifers. Twenty-four Braford heifers were randomly assigned to four diets where sorghum silage was replaced at 0%, 30%, 60%, and 100% for triticale silage in a feedlot system. During sixty-tree days of the experiment, the feed intake, feed ratio conversion, and average daily gain were measured. The heifers were slaughtered and the effects of the sorghum and triticale silage in carcass characteristics were evaluated. The replacement of sorghum silage with triticale silage did not affect the feed intake and average daily gain (P> 0.05). Feed gain ratio was higher for the heifers that received lower proportions of triticale silage (0% and 30%) in replace sorghum silage (P <0.05). Eye loin area and fat thickness were similar among treatments (P<0.05). Other carcass characteristics, such as slaughter live weight, dressing percentage, conformation and fat classification were similar among experimental treatments (P<0.005). Triticale silage can replace sorghum silage for finishing beef heifers in feedlot system and provides similar carcass characteristics.(AU)

O objetivo deste trabalho foi avaliar o efeito da substituição da silagem de sorgo por diferentes proporções de silagem de triticale no desempenho e na carcaça de novilhas Braford. A silagem de sorgo foi substituída em 0%, 30%, 60% e 100% pela silagem de triticale como fonte de volumoso na alimentação de 24 novilhas da raça Braford, em um sistema de confinamento. O período experimental total foi de 63 dias para a avaliação do consumo dos componentes da ração, a conversão alimentar e o ganho médio diário. Ao final do experimento, os animais foram abatidos e avaliados em relação às características de carcaça. A substituição da silagem de sorgo pela silagem de triticale não apresentou efeito no consumo dos componentes da ração e no ganho médio diário entre as novilhas (P>0,05). A conversão alimentar foi melhor para as novilhas que receberam menores proporções de silagem de triticale (0% e 30%) em substituição à silagem de sorgo (P<0,05). As características das carcaças foram semelhantes entre os animais independentemente da fonte de volumoso. A silagem de triticale apresenta-se como alimento alternativo à silagem de sorgo em regiões de transição climática para a terminação de novilhas de corte, proporcionando o mesmo desempenho animal e as mesmas características de carcaça.(AU)

Estabilidade genética em triticale estimada pela viabilidade polínica / Genetic stability in triticale estimated by pollen viability

O estudo visou inferir sobre a estabilidade genética, com base na viabilidade polínica, em genótipos de triticale hexaploide utilizados no bloco de cruzamentos do programa de melhoramento genético da Empresa Brasileira de Pesquisa Agropecuária (Embrapa Trigo). O experimento foi conduzido em delineamento inteiramente casualizado. Para cada genótipo, foram avaliadas cinco repetições e cada repetição foi constituída por uma planta. Analisaram-se, por meio de microscopia óptica e pela técnica de Squash com corante carmim acético 1%, 200 grãos de pólen por lâmina, totalizando 1.000 grãos de pólen por genótipo. As variáveis analisadas foram: grãos de pólen binucleados e trinucleados, com pouco amido, vazios, com mais de um poro e de tamanhos diferentes. Os dados obtidos foram submetidos à análise de variância e as médias comparadas pelo teste de Tukey a 1%. Houve diferença significativa entre todas as variáveis. A porcentagem de grãos de pólen binucleados e/ou trinucleados variou de 74 a 97%. Concluiu-se que 66% dos genótipos avaliados apresentaram viabilidade polínica acima de 90%, sendo indicados a continuar fazendo parte do programa de melhoramento genético de triticale, tanto na seleção de parentais como durante os cruzamentos e retrocruzamentos. Portanto, os estudos citogenéticos representam excelente ferramenta de apoio ao melhorista na escolha dos genótipos mais estáveis.(AU)

The study aimed to infer about the genetic stability from the pollen viability in hexaploid triticale genotypes used in block crossings breeding program from Brazilian Company of Agriculture Research (Embrapa Wheat). The experiments were conducted in a randomized design. For each genotype five repetitions were assessed. Each replication consisted of one plant and each replication consisted of one plant. Two hundred pollen grains were analyzed by optical microscopy and by the squash technique with 1% acetic carmine dye per slide, totalizing 1,000 pollen grains per genotype. The variables analyzed were: binucleate and trinucleate pollen grains, with little starch, empty, with more than one pore and with different sizes. The data were submitted to analysis of variance and compared by Tukey test at 1%. There were significant differences between all variables. The percentage of binucleate and/or trinucleate grains ranged from 74 to 97%. We conclude that 66% of genotypes present pollen viability above 90% and they are indicated to remain of the breeding program of triticale, both in the selection of parenting as during the crossing and backcrossing. Therefore, the cytogenetic studies represent excellent support tool for breeder in selecting the most stable genotypes.(AU)

Avaliação de linhagens de triticale na região do Alto Paranaíba ­ MG / Evaluation of triticale inbred lines in the region of Alto Paranaíba, state of Minas Gerais, Brazil

O presente trabalho tem por objetivo avaliar novas linhagens de triticale, em dois locais do Estado de Minas Gerais, visando à seleção de novos genótipos para serem multiplicados e distribuídos aos agricultores. Os experimentos foram constituídos de 13 linhagens de triticale e três cultivares de trigo, em delineamento estatístico de blocos ao acaso com três repetiçoes. Foi avaliado o rendimento de grãos e o peso hectolitro. As análises de variância individuais e conjunta demonstraram efeitos significativos de genótipos para as características avaliadas e significativos quando considerado genótipo x local. Todas as linhagens de triticale apresentaram altas produtividades, bem maiores que as obtidas pelas cultivares de trigo. As altas produtividades destas linhagens de triticale demonstram o alto potencial destes genótipos para ser avaliados em futuros ensaios de competição, para o lançamento de futuras cultivares, com destaque, considerando as variáveis avaliadas, para as linhagens EP 068009, EP 068012, EP 068055 e EP 068050, nos dois locais cultivados. O peso do hectolitro das linhagens de triticale foi baixo (média igual a 73,5 kg/hl), o que pode ser explicado pelo comprimento dos grãos e a sua forma, além do fator genético resultante do cruzamento trigo x centeio.

The objective of this work was to evaluate the new breeds lines of triticale in the regions of Minas Gerais state aiming the selection of new germoplasma to be multiplied and distributed among agricultors. The experiments were constituted by 13 lines of triticale and 3 cultivars of wheat ramdomly in block. The grains growth and produce and the yield hectoliter weight, were evaluated. The analizes of individual and group variations presented highly significant variation. All lines presented high productivity, greater than the ones cultivated by wheat. The high productivity of the triticale lines demostrate the material great potential to be analized in future competitional experiments envisioning the future cultivational methods to be launched, outlining the EP 068009, EP 068012, EP 068050 e EP 068055 genotypes in both cultivated ambient. The yield hectoliter weight, of the triticale lines were very low, what can be explained by the grain length and shape, besides the genetic factor as the result of wheat x rye.