Добірка наукової літератури з теми "Gene Gpc-B1 from Triticum turgidum ssp. dicoccoides"

Aim. The purpose of our study was to determine the content of total protein in the F5 generation grains, the carriers of the Gpc-B1 gene from Triticum turgidum ssp. dicoccoides by two methods, which in turn would confirm the successful expression of the Gpc-B1 gene in the genetic environment of bread winter wheat. Methods. Determination of protein content was carried out by Kjeldahl method and by infrared spectrometry (NIR) method. Results. The 44 hybrid lines that are homozygous for the Gpc-B1 gene from T. turgidum ssp. dicoccoides have been analyzed. It has been established that for both methods, the average content of protein in the grain of hybrid lines is 14 % higher in comparison to the original Kuyalnik variety. Particular attention should be paid to the line number 10, 12 and 35 in which the content of protein exceeds 15 % by the method of Kjeldahl. Conclusions. The obtained results indicate that the gene Gpc-B1 from the wild relative in the new genetic environment of the highly productive registered wheat cultivar Kuyalnik has been functioning and has a positive effect on the accumulation of total protein in grains.Keywords: biofortification, protein content, Triticum aestivum, Gpc-B1 gene, Kjeldahl and NIR methods.

The aim of this study was to investigate the effect of zinc (Zn) deficiency on the growth and grain yield of wheat with different allele statuses of the Gpc-B1 gene. For this research, common wild emmer wheat (Triticum turgidum ssp. dicoccoides (Koern. ex Asch. &Graebn.) Schweinf.), bread wheat (Triticum aestivum L. cv. Festivalnaya), and two intogressive lines were used. T. dicoccoides and introgressive line 15-7-1 carry a functional allele of the Gpc-B1 gene, while the T. aestivum cv. Festivalnaya and introgressive line 15-7-2 carry the non-functional Gpc-B1 allele. Zn deficiency did not affect the shoot height or fresh weight of any of the studied plants. The only exception was T. dicoccoides, where a small decrease in shoot height was registered. Additionally, under Zn deficiency T. dicoccoides had an increase in flag leaf area, spike length, and dry weight, as well as in grain number and grain yield per spike. The other variants did not experience changes in the above-described parameters under Zn deficiency. Under Zn deficiency, the Zn concentration in the grains was higher in the plants with a functional allele of the Gpc-B1 gene compared to the plants with a non-functional allele. These results show that wheat with a functional allele of the Gpc-B1 gene growing under Zn deficiency is capable of grain production with a sufficient Zn concentration without a decrease in yield.

The high cost of cultivar development encourages efficiencies to reduce time and costs to develop cultivars. Doubled haploid (DH) technology and marker assisted breeding (MAB) are two such tools that improve efficiencies. Since 1997, twenty five wheat cultivars in seven market classes, developed using DH methods, have been registered by the Canadian Food Inspection Agency. These DH cultivars accounted for more than one third of the Canadian wheat acreage in 2009. The DH cultivar Lillian, eligible for grades of Canada Western Red Spring class and currently the most widely grown wheat cultivar in Canada, was developed using MAB to improve grain protein content with the Gpc-B1/Yr36 on chromosome 6BS introgressed from Triticum turgidum L. (Zhuk.) dicoccoides (Körn. Ex Asch. & Graebn). AC Andrew, a Canada Western Soft White spring DH cultivar, was the most widely grown cultivar in its class for the last two years. The new market class, Canada Western Hard White Spring wheat, is based entirely on DH cultivars. Goodeve, one of the first Canada Western Red Spring cultivars released with the gene Sm1 on chromosome 2BS for resistance to the orange wheat blossom midge (Sitodiplosis mosellana (Géhin)) was selected by application of the DNA marker WM1. Glencross, the first cultivar in the Canada Western Extra Strong wheat class with Sm1, was selected using the WM1 marker on haploid plants prior to doubling. Development of the durum wheat cultivars CDC Verona and Brigade involved the use of a marker for Cdu1, a major gene on chromosome 5B that regulates grain cadmium concentration. Marker technology permits a more strategic and integrated approach to breeding by quantifying the introgression of various key genes into advanced breeding material, identifying targeted loci in parents and following up with MAB in the progeny.

Дисертаційна робота присвячена біотехнологічним аспектам біофортифікації озимої м’якої пшениці геном Gpc-B1 від Тriticum turgidum ssp. dicoccoides, що підвищує вміст білка та мінеральних елементів. Розроблено молекулярно генетичні маркерні системи для комплексного аналізу та відбору рослин з популяції м’якої озимої пшениці, носіїв гена Gpc-B1 від Тriticum turgidum ssp. dicoccoides. З 160-ти дослідних сімей F2 покоління, було відібрано 44-и ліній F5 покоління, гомозиготних за цільовим геном. Вони були комплексно охарактеризовані по генам пуроіндолінів та глютенінів, проведено вимірювання вмісту макро- та мікроелементів, визначено вміст загального білка у зерні та зрештою відібрано 13-ть ліній, які є цінним і перспективним матеріалом для подальших селекційних робіт. Проведено аналіз врожайності та фізіологічних показників ліній пшениці, носіїв гена Gpc-B1 від Triticum turgidum ssp. dicoccoides, який показав, що лінії є стійкими до полягання, середньорослими з середньою врожайністю, високим вмістом та якістю запасного білка.
The dissertation is devoted to the biotechnological aspects of winter bread wheat biofortification by the Gpc-B1 gene introgressed from Triticum turgidum ssp. dicoccoides, which increases the content of protein and minerals in grain. Molecular-genetic marker systems for the complex analysis and selection of bread wheat hybrids, the carriers of Gpc-B1 gene from Triticum turgidum ssp dicoccoides, have been developed. The 44 lines of F5 generation, homozygous in target gene were selected from 160 plants of F2 generation. The generation of F5 hybrid lines was thoroughly characterized by the puroindolins and glutenins genes. The macro- and micronutrient content analysis was performed. The content of total protein in grain was determined, and eventually, 13 hybrid lines were selected for the future work. The analysis of yield and physiological parameters of lines-carriers of Gpc-B1 gene from Triticum turgidum ssp. dicoccoides showed that the lines were resilient to lodging, medium-growing with medium yields.
Диссертация посвящена биотехнологическим аспектам биофортификации озимой мягкой пшеницы геном Gpc-B1 от Тriticum turgidum ssp. dicoccoides, который повышает содержание белка и минеральных элементов в зерне. Разработаны молекулярно генетические маркерные системы для комплексного анализа и отбора образцов мягкой пшеницы, носителей гена Gpc-B1 от Тriticum turgidum ssp. dicoccoides. С их помощью для SSR локусов Xgwm626, Xgwm508, Xgwm193, Xgwm219, расположенных на 6В хромосоме, было рассчитано частоту рекомбинации, которая составила для Xgwm508 = 2,94 ± 1,28%, Xgwm193 = 3,96 ± 1,51%, Xgwm626 = 2,98 ± 1,29%, Xgwm219 = 6,77 ± 2,08%. Из 160-ти семей F2 поколения, методическим ежегодным анализом было отобрано 44 линии F5 поколения, гомозиготных по гену Gpc-B1, перенесенным из Тriticum turgidum ssp. dicoccoides. Проведен анализ аллельного состояния пуроиндолинових генов F5 поколения линий пшеницы, носителей гена Gpc-B1 от Triticum turgidum ssp. dicoccoides, а также был измерен физический показатель твердозерности. Выявлено 17 линий, имеющих такое же аллельное состояние генов, как у исходного сорта Куяльник, а также обнаружены уникальные генотипы, несущие комбинацию Pina-D1b, Pinb-D1a, которую не было выявлено ранее в сортах украинской селекции. Статистическая обработка данных показала, что сочетание аллелей Pina-D1b Pinb-D1a статистически достоверно (р<0,05) обеспечивало большую твердозерность, чем у растений, с аллеями Pina-D1a Pinb-D1b. В ходе выполнения работы был проведен анализ аллельного состояния генов глютенинов озимых линий пшеницы F5 поколения и отобрано 16-ть наиболее ценных, содержащих оптимальную для хлебопекарского качества аллельную формулу локуса Glu-1 (Glu-A1a или Glu-A1b, Glu-B1al, Glu-D1d). Проанализированы линии пшеницы F4 и F5 поколений на содержание макро- и микроэлементов в зерне методом ICP-MS. Было установлено, что присутствие гена Gpc-B1 от Triticum turgidum ssp. dicoccoides в линиях пшеницы приводит к статистически (на уровне 0,05) значимому повышению уровня накопления биологически важных элементов питания – железа, цинка, марганца, меди, селена, а также магния. Проведен комплексный анализ определения содержания общего белка в зерне растений F5 поколения методом Кьельдаля и NIR. В среднем содержание белка в линиях, которые были носителями гена Gpc-B1 из Triticum turgidum ssp. dicoccoides, повышалось на 14% по сравнению с исходным сортом Куяльник. В результате было отобрано 13-ть линий, перспективных для дальнейшей работы. Разработанная технология отбора по урожайности и физиологическим показателям позволила в F6 поколении носителей гена Gpc-B1 от Triticum turgidum ssp. dicoccoides определить линии устойчивые к полеганию, среднерослые и со средней урожайностью, которая даже превышала среднюю урожайность сортов озимой пшеницы 2017 года в Киевской области, и на была 40% выше чем у исходной линии Glupro. Нами был проведен анализ хлебопекарного качества зерна пшеницы методом непрямой оценки «силы» муки – индексом седиментации SDS-30. У большинства линий пшеницы он был больше 80 мл, что считается очень хорошим показателем. Высокий индекс седиментации SDS-30, указывает на отменное хлебопекарное качество муки. Суммируя все показатели разработанной технологии отбора можно отметить, что линии-носители гена Gpc-B1 от Triticum turgidum ssp. dicoccoides являются уникальным генетическим материалом, который сочетает в себе лучшие свойства родителей и в дальнейшем будет использоваться для создания новых высокоперспективных сортов пшеницы.

High Grain Protein Content (GPC) of wheat is important for improved nutritional value and industrial quality. However, selection for this trait is limited by our poor understanding of the genes involved in the accumulation of protein in the grain. A gene with a large effect on GPC was detected on the short arm of chromosome 6B in a Triticum turgidum ssp. dicoccoides accession from Israel (DIC, hereafter). During the previous BARD project we constructed a half-million clones Bacterial Artificial Chromosome (BAC) library of tetraploid wheat including the high GPC allele from DIC and mapped the GPC-B1 locus within a 0.3-cM interval. Our long-term goal is to provide a better understanding of the genes controlling grain protein content in wheat. The specific objectives of the current project were to: (1) complete the positional cloning of the GPC-B1 candidate gene; (2) characterize the allelic variation and (3) expression profile of the candidate gene; and (4) validate this gene by using a transgenic RNAi approach to reduce the GPC transcript levels. To achieve these goals we constructed a 245-kb physical map of the GPC-B1 region. Tetraploid and hexaploid wheat lines carrying this 245-kb DIC segment showed delayed senescence and increased GPC and grain micronutrients. The complete sequencing of this region revealed five genes. A high-resolution genetic map, based on approximately 9,000 gametes and new molecular markers enabled us to delimit the GPC-B1 locus to a 7.4-kb region. Complete linkage of the 7.4-kb region with earlier senescence and increase in GPC, Zn, and Fe concentrations in the grain suggested that GPC-B1 is a single gene with multiple pleiotropic effects. The annotation of this 7.4-kb region identified a single gene, encoding a NAC transcription factor, designated as NAM-B1. Allelic variation studies demonstrated that the ancestral wild wheat allele encodes a functional NAC transcription factor whereas modern wheat varieties carry a non-functional NAM-B1 allele. Quantitative PCR showed that transcript levels for the multiple NAMhomologues were low in flag leaves prior to anthesis, after which their levels increased significantly towards grain maturity. Reduction in RNA levels of the multiple NAMhomologues by RNA interference delayed senescence by over three weeks and reduced wheat grain protein, Zn, and Fe content by over 30%. In the transgenic RNAi plants, residual N, Zn and Fe in the dry leaves was significantly higher than in the control plants, confirming a more efficient nutrient remobilization in the presence of higher levels of GPC. The multiple pleiotropic effects of NAM genes suggest a central role for these genes as transcriptional regulators of multiple processes during leaf senescence, including nutrient remobilization to the developing grain. The cloning of GPC-B1 provides a direct link between the regulation of senescence and nutrient remobilization and an entry point to characterize the genes regulating these two processes. This may contribute to their more efficient manipulation in crops and translate into food with enhanced nutritional value. The characterization of the GPC-B1 gene will have a significant impact on wheat production in many regions of the world and will open the door for the identification of additional genes involved in the accumulation of protein in the grain.

High Grain Protein Content (GPC) is a desirable trait in breadmaking and pasta wheat varieties because of its positive effects on quality and nutritional value. However, selection for GPC is limited by our poor understanding of the genes involved in the accumulation of protein in the grain. The long-term goal of this project is to provide a better understanding of the genes controlling GPC in wheat. The specific objectives of this project were: a) to develop a high-density genetic map of the GPC gene in tetraploid wheat, b) to construct a T. turgidum Bacterial Artificial Chromosome (BAC) library, c) to construct a physical map of the GPC gene and identify a candidate for the GPC gene. A gene with a large effect on GPC was detected in Triticum turgidum var. dicoccoides and was previously mapped in the short arm of chromosome 6B. To define better the position of the Gpc-B1 locus we developed homozygous recombinant lines with recombination events within the QTL region. Except for the 30-cM region of the QTL these RSLs were isogenic for the rest of the genome minimizing the genetic variability. To minimize the environmental variability the RSLs were characterized using 10 replications in field experiments organized in a Randomized Complete Block Design, which were repeated three times. Using this strategy, we were able to map this QTL as a single Mendelian locus (Gpc-B1) on a 2.6-cM region flanked by RFLP markers Xcdo365 and Xucw67. All three experiments showed that the lines carrying the DIC allele had an average absolute increase in GPC of 14 g/kg. Using the RFLP flanking markers, we established the microcolinearity between a 2.l-cM region including the Gpc-B1 gene in wheat chromosome 6BS and a 350-kb region on rice chromosome 2. Rice genes from this region were used to screen the Triticeae EST collection, and these ESTs were used to saturate the Gpc-B1 region with molecular markers. With these new markers we were able to map the Gpc-B1 locus within a 0.3-cM region flanked by PCR markers Xucw83 and Xucw71. These flanking markers defined a 36-kb colinear region with rice, including one gene that is a potential candidate for the Gpc-B1 gene. To develop a physical map of the Gpc-B1 region in wheat we first constructed a BAC library of tetraploid wheat, from RSL#65 including the high Gpc-B1 allele. We generated half- million clones with an average size of l3l-kb (5.1 X genome equivalents for each of the two genomes). This coverage provides a 99.4% probability of recovering any gene from durum wheat. We used the Gpc-BI flanking markers to screen this BAC library and then completed the physical map by chromosome walking. The physical map included two overlapping BACs covering a region of approximately 250-kb, including two flanking markers and the Gpc-B1 gene. Efforts are underway to sequence these two BACs to determine if additional wheat genes are present in this region. Weare also developing new RSLs to further dissect this region. We developed PCR markers for flanking loci Xucw79andXucw71 to facilitate the introgression of this gene in commercial varieties by marker assisted selection (httQ://maswheat.ucdavis.edu/ orotocols/HGPC/index.hlm). Using these markers we introgressed the Gpc-B1 gene in numerous pasta and common wheat breeding lines.