Experssion vectors for editing kafirin (k1C5, gKAF1) and ameiotic 1 genes in sorghum by agrobacterial transformation

Biomics ◽  
2020 ◽  
Vol 12 (4) ◽  
pp. 520-531
Author(s):  
G.A. Gerashchenkov ◽  
L.A. Elkonin ◽  
N.A. Rozhnova

One of the effective approach to improve the nutritional value of sorghum grains is the production of mutants with partially or completely blocked synthesis or the altered amino acid composition of spare proteins - kafirins. The usage of the genomic editing method allows to solve this problem by introducing mutations into the nucleotide sequences of the genes α- and γ-kafirins (k1C5 and gKAF1). Another important area in the genetics and selection of agricultural plants is the development and use of apomixis technologies to fix heterosis in F1 hybrids. The DYAD arabidopsis gene and its homolog ameiotic 1 are the potential apomixis candidate genes involved in the normal course of meiosis, the induction mutations in which is one of the way to solve this problem. In the course of research, the genomic targets (20 bp sequences) were selected to introduce mutations into the genes of α- and γ-kafirins of sorghum and ameiotic 1 gene through CRISPR/Cas genomic editing technology. Guide RNA design is performed using bioinformatic programs CRISPROR and CHOPCHOP. Cloning of the α- and γ- kafirins and ameiotic 1 targets of sorghum into vector pBAtC was carried out at sites AarI. Validation of the success of target cloning was performed by DNA sequencing. The created constructions were transferred by electroporation to the agrobacterial strain AGL0. Currently, the created vectors are used for the agrobacterial transformation of sorghum plants.

Author(s):  
O. V. Gladysheva ◽  
Т. А. Barkovskaya

This article presents the results of a study of hybrid spring wheat populations. Revealed different patterns of inheritance of important traits in F1 hybrids, which is caused by hereditary features of the original forms and circumstances. Analysis of hybrid populations showed that the dominant role in the formation of 1000 grains mass belongs to productivity and weight of grain with an ear, correlation coefficient r = 0.90 and amounted to r = 0.73, respectively. Found that the hybrid population in F4-5, created on the basis of grades Moscow 35, Esther, Agatha, Rome, Saratovskaya 29 (Russia), Ostinka (Ukraine) are a valuable material for selection of highly productive genotypes for future use breeding process. 


Gene Therapy ◽  
2021 ◽  
Author(s):  
Jonathan O’Keeffe Ahern ◽  
Irene Lara-Sáez ◽  
Dezhong Zhou ◽  
Rodolfo Murillas ◽  
Jose Bonafont ◽  
...  

AbstractRecent advances in molecular biology have led to the CRISPR revolution, but the lack of an efficient and safe delivery system into cells and tissues continues to hinder clinical translation of CRISPR approaches. Polymeric vectors offer an attractive alternative to viruses as delivery vectors due to their large packaging capacity and safety profile. In this paper, we have demonstrated the potential use of a highly branched poly(β-amino ester) polymer, HPAE-EB, to enable genomic editing via CRISPRCas9-targeted genomic excision of exon 80 in the COL7A1 gene, through a dual-guide RNA sequence system. The biophysical properties of HPAE-EB were screened in a human embryonic 293 cell line (HEK293), to elucidate optimal conditions for efficient and cytocompatible delivery of a DNA construct encoding Cas9 along with two RNA guides, obtaining 15–20% target genomic excision. When translated to human recessive dystrophic epidermolysis bullosa (RDEB) keratinocytes, transfection efficiency and targeted genomic excision dropped. However, upon delivery of CRISPR–Cas9 as a ribonucleoprotein complex, targeted genomic deletion of exon 80 was increased to over 40%. Our study provides renewed perspective for the further development of polymer delivery systems for application in the gene editing field in general, and specifically for the treatment of RDEB.


2021 ◽  
pp. 34-37
Author(s):  
Р.Х. Беков

Рассмотрены важные аспекты использования геноносителей различных маркерных признаков при создании и селекции гетерозисных гибридов томата для защищенного грунта. Наряду с общими требованиями к родительским компонентам, касающимися продуктивности растений, скороспелости, устойчивости к болезням и т. д., сегодня при селекции на гетерозис для защищенного грунта к исходному материалу предъявляют более высокие требования по наличию хозяйственно полезных признаков. Эти требования касаются типа роста растений, формы, окраски и качества плодов, типа кисти, формы плодоножки и т. д. Представлены характеристики исходного материала для создания гетерозисных гибридов томата, а также даны характеристики полученных гибридов. Исследования, проведенные в отделе селекции ВНИИО (ВНИИ овощеводства – филиал ФГБНУ ФНЦО), показали большую перспективность использования при гетерозисной селекции томата для защищенного грунта геноносителей различных маркерных признаков, особенно признаков, определяющих коричневую окраску эндосперма семян (гены bs и bs-2) и несочлененную плодоножку (ген j-2). Созданные с использованием этих маркерных признаков гибриды и селекционные линии по-своему уникальны и являются приоритетным достижением этого научного учреждения, так как они представляют большой интерес для гетерозисной селекции томата в качестве исходного материала. Целый ряд гибридов F1 включен в Госреестр селекционных достижений, допущенных к использованию (Удача, Голубчик, Клад овощевода, Заур и др.). В связи с развитием фермерского и любительского овощеводства в последние годы определенное внимание было уделено созданию новых мутантных линий с розовыми, малиновыми и темно-коричневыми плодами с высокими вкусовыми качествами (учитывая, что образцы такого типа пользуются повышенным спросом у населения и стоят дороже). При создании новых гетерозисных гибридов были учтены эти признаки родительских форм. Important aspects of the use of gene carriers of various marker traits in the creation and selection of heterotic tomato hybrids for greenhouse industry are considered. Along with the general requirements for the parent components concerning plant productivity, precocity, resistance to diseases, etc., today, when breeding for heterosis for greenhouse industry, higher requirements are imposed on the source material for the presence of economically useful features. These requirements relate to the type of plant growth, shape, colour and quality of fruits, brush type, stem shape, etc. The characteristics of the initial material for creating heterotic tomato hybrids are presented, as well as the characteristics of the resulting hybrids are given. Studies conducted in the breeding department of the ARRIVG (ARRIVG – a branch of the FSBI FSCV) showed great prospects for the use of gene carriers of various marker traits, especially those that determine the brown colour of the seed endosperm (bs and bs-2 genes) and an undifferentiated peduncle (j-2 gene) in the heterosis breeding of tomatoes for greenhouses. Hybrids and breeding lines created using these marker traits are unique in their own way and are a priority achievement of this scientific institution, since they are of great interest for the heterosis breeding of tomato as a basic material. A number of F1 hybrids are included in the State Register of Breeding Achievements allowed for use (Udacha, Golubchik, Klad ovoschevoda, Zaur, etc.). Due to the development of farming and amateur vegetable growing in recent years, some attention has been paid to the creation of new mutant lines with pink, crimson and dark brown fruits with high taste qualities (given that samples of this type are in high demand among the population and are more expensive). When creating new heterotic hybrids, these features of the parent forms were taken into account.


2015 ◽  
Vol 33 (8) ◽  
pp. 805-806 ◽  
Author(s):  
Cameron Ross MacPherson ◽  
Artur Scherf
Keyword(s):  

Author(s):  
G.F. Monakhos ◽  
S.G. Monakhos ◽  
R.R. Alizhanova

На Селекционной станции имени Н.Н. Тимофеева гибридизацией с донором устойчивости к пероноспорозу F1 Santero, беккроссом, инбридингом и отбором на инфекционном фоне с помощью молекулярного маркера DMR1 создано 15 линий лука репчатого гомозиготного по гену устойчивости Pd1. Оценка в однолетней культуре позволила выделить линии с массой луковиц 100120 г с высокой сохранностью, которые могут быть использованы в селекции F1 гибридов в качестве отцовских компонентов.Fifteen homozygous for Pd1 downy mildew resistance gene onion lines were developed by hybridization followed backcrossing F1 Santero of downy mildew resistant onion accession of European origin and a collection of onion inbred lines from LC Breeding station after N.N. Timofeev. The resistant to downy mildew, caused by Peronospora destructor (Berk.), plants were selected in segregated populations based on disease resistance test and molecular genotyping using DMR1marker. The lines with a mass of bulbs 100 120 g with high preservation, which can be used in the selection of F1 hybrids as paternal components.


Author(s):  
V.M. Lukomets ◽  
◽  
S.V. Zelentsov ◽  
E.V. Moshnenko ◽  
◽  
...  

Breeding practice shows that soybean cultivars developed by synthetic breeding methods are submitted for the state variety testing in the F10–F11 generation. But the newly bred cultivars are not completely homozygous. The studies were related to the determination of the frequencies of formation of atypical and suitable for selection promising individuals in soybean cultivars developed by synthetic breeding methods. The studies were carried out in 2019–2020 at the central experimental base of V.S. Pustovoit All-Russian Research Institute of Oil Crops, Krasnodar. In the experiments, we used cultivars of our own breeding: Selena, Puma, Vita, Irbis, Bars, Barguzin and Sayana with a total age of 11–15 years from the year F1 hybrids were obtained. To confirm the practical possibility of isolating individuals differing from the phenotypic varietal norm in varietal populations, a complete examination of the crops of all studied soybean cultivars was carried out. In the fields of all cultivars, individuals were identified that differed from the varietal norm phenotypically. Mostly, the isolated individuals were distinguished by an increased plant height, a more powerful habit, an increased number of beans per plant, and resistance to lodging. The facts of the detection of atypical individuals with improved morphological traits in soybean cultivars of different ages confirm the possibility of individual selection of plants in varietal populations based on morphometric traits. The statistical dynamics of a decrease in heterozygous individuals in increasing generations in a hybrid self-pollinating population in terms of the number of paired genes, by which the parental forms can hypothetically differ, were calculated using the modified formula of S. Borojević (1984). It was found that the frequency of formation of phenotypically different individuals in varietal populations of soybeans depends on the total age of the cultivar. The frequency of the formation of morphologically different individuals decreases with an increase in the number of generations of the cultivar. Individual selection of individuals with positive phenotypic differences from the varietal norm can be recommended as an additional source of promising and practically homozygous starting material for accelerated analytical breeding of soybean.


2022 ◽  
Vol 8 ◽  
Author(s):  
María D. Raigón ◽  
María D. García-Martínez ◽  
Octavian P. Chiriac

The loss of genetic diversity due to the replacement of local tomato (Solanum lycopersicum L.) varieties by improved cultivars has been mitigated in many cases by the good work of organic farmers in maintaining local agricultural biodiversity. In parallel to these initiatives, in recent years, consumers have developed an increasing awareness of both food-related health, environmental issues, and food demand to recover the flavors of the past. In the case of tomatoes, these attributes (nutritional, organoleptic, social, and environmental) are closely related to organic production using local varieties. “Malacara” tomato is an example of a local variety. Coming from Sierra de Cádiz, it is a varietal type called “Cuelga” (“for hanging,” because the tomato trusses are hung from beams in the farmhouses). Cultivated and harvested in the open air during the summer months, these tomatoes are commercialized and consumed in the winter. Historically, this variety has enabled the fresh consumption of tomatoes during the winter, without the need to force cultivation. It is highly appreciated in the local cuisine and is the basis for sauces figuring in typical dishes. Its characteristic traits are small, pallid fruits, and long shelf life. The main objective of this work has been to typify two Malacara tomato cultivars (red and yellow color) grown under organic farming conditions, through the characterization of morphological, nutritional, and volatile parameters. The main differences are due to morphological parameters (fruit weight and color of the exocarp and endocarp). Other characteristics such as the content of ash, fiber, moisture, the concentration of iron, magnesium, and calcium, and content of lycopene are different between both cultivars. This study provides information on the nutritional and aromatic composition of two Malacara tomato cultivars, differentiated by their color and grown under organic farming conditions. The results add value to the native horticultural heritage and can aid in the selection of tomato varieties suitable for a sustainable production system and to produce tomatoes with high nutritional value and rich in aroma.


Author(s):  
N.V. Geraskina ◽  
V.V. Ognev

Представлены актуальные направления селекции баклажана для условий открытого и защищенного грунта. Выделен перспективный исходный материал для различных направлений использования. Дана характеристика перспективным образцам для использования в селекции. В качестве исходного материала для селекции целесообразно использовать сорта и гибриды F1 Эскимо, Халиф, Меч самурая Десерт Голиафа, Умка, Альбион и новый линейный материал Кит 53, Л 67, Л 83, 85/17, 94/16, Л 107.The perspective directions of selection of eggplant for conditions of open and protected soil are presented. The perspective source material for different directions of use is allocated. The characteristic of perspective samples for use in selection is given. As initial material for breeding it is advisable to use the obtained varieties and F1 hybrids Eskimo, Caliph, Sword of the samurai, Goliath Dessert, Umka, Albion, new line material Kit 53, L 67, L 83, 85/17, 94/16, L 107.


Proceedings ◽  
2020 ◽  
Vol 53 (1) ◽  
pp. 14
Author(s):  
Francisco Teodoro Rios ◽  
Argentina Angelica Amaya ◽  
Manuel Oscar Lobo ◽  
Norma Cristina Samman

The aim was to develop a snack bar using regional food products. The formulation included traditional cereals and amaranth, quinoa, sunflower, flax, chia, sesame and poppy seeds subjected to different treatments. Two sensory evaluations were carried out to evaluate acceptability. Snack bars containing toasted seeds presented high acceptability by the consumer. Amaranth, quinoa, chia and sunflower significantly increased the acceptability. The sensory methods applied allowed for the selection of ingredients and processing technologies that increase the preference of consumers.


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