Somatic hybrids between potato Solanum tuberosum and wild species Solanum verneï exhibit a recombination in the plastome

2005 ◽  
Vol 83 (1) ◽  
pp. 1-11 ◽  
Author(s):  
S. Trabelsi ◽  
R. Gargouri-Bouzid ◽  
F. Vedel ◽  
A. Nato ◽  
L. Lakhoua ◽  
...  
Keyword(s):  
Solanum Tuberosum ◽  
Wild Species ◽  
Somatic Hybrids ◽  
Solanum Vernei

Разработка и тестирование универсальных праймеров для ПЦР-амплификации генов-ортологов β-фруктофуранозидазы (Pain-1) у видов и сортов картофеля

2018 ◽  
Author(s):  
E.O. Shmelkova ◽  
M.A. Slugina ◽  
A.A. Meleshin ◽  
E.V. Romanova
Keyword(s):  
Solanum Tuberosum ◽  
Wild Species ◽  
Starch Synthesis ◽  
Full Length ◽  
Universal Primers ◽  
Potato Tubers ◽  
Vacuolar Invertase ◽  
Reserve Carbohydrate ◽  
Cold Induced Sweetening ◽  
Design And Testing

Работа посвящена разработке и тестированию универсальных праймеров для ПЦР-амплификации полноразмерных генов-ортологов β-фруктофуранозидазы (кислой вакуолярной инвертазы) у видов и сортов картофеля (Solanum tuberosum). Крахмал – основной источник энергии и резервный углевод, накапливающийся в амилопластах клубней. Образовавшаяся в результате фотосинтеза молекула глюкозы при реакции с фруктозой образует сахарозу – основную транспортную форму углеводов в растении. В клубни сахароза доставляется по флоэме (апопластный путь), где в межклеточном пространстве расщепляется до глюкозы и фруктозы, которые затем проникают в клетки паренхимы. Глюкоза служит в дальнейшем субстратом для синтеза крахмала в амилопластах. Однако при воздействии пониженных температур крахмал в клубнях картофеля разрушается до редуцирующих сахаров. Параллельно этому процессу идет ресинтез сахарозы до глюкозы и фруктозы за счет фермента кислой вакуолярной инвертазы (β-фруктофуранозидазы), кодируемой геном Pain-1. В совокупности эти процессы приводят к избыточному накоплению моносахаров в клубнях картофеля, так называемому холодовому осахариванию (cold-induced sweetening). При этом создаются условия для интенсивного образования меланоидинов, вызывающих потемнение мякоти картофеля, что значительно ухудшает товарное качество продукта. Таким образом, изучение гена Pain-1, кодирующего вакуолярную инвертазу, а именно, его идентификация и анализ структуры – важная задача, необходимая для поиска доноров, устойчивых к холодовому осахариванию. Первоочередная задача для этого – разработка и тестирование праймерных комбинаций, позволяющих амплифицировать полноразмерный ген у диких видов картофеля, а также сортов и линий культивируемого картофеля (S. tuberosum). В данной работе приведены результаты разработки и тестирования универсальных праймеров, с помощью которых можно амплифицировать как полноразмерные гены-ортологи, так и фрагменты гена Pain-1, а также подобраны оптимальные условия для проведения ПЦР реакции. Было разработано 6 праймерных комбинаций (PainF – PainR, PainF – Pain1exR, Pain1exF – Pain3exR, Pain2inF – Pain2inR, Pain3exF – Pain5exR, Pain5exF – PainR), среди которых комбинация PainF – PainR позволяла амплифицировать полноразмерный ген, остальные – внутренние и будут использованы в дальнейшем при секвенировании фрагментов исследуемого гена. Эти праймеры были успешно протестированы на 15 образцах, включающих представителей пяти дикорастущих видов картофеля (S. gourlay, S. chacoense, S. pinnatissectum, S. stoloniferum, S. vernei) и десяти сортов российской и зарубежной селекции (Гала, Ласунок, Ред Скарлетт, Рассет Бербанк, Мирас, Башкирский, Жуковский ранний, Матушка, Елизавета, Сударыня).The purpose of research is design and testing of universal primers for PCR amplification of full-length-fructofuranozidase orthologs genes (acid vacuolar invertase) in wild species and potato (Solanum tuberosum) varieties. Starch is the main source of energy and a reserve carbohydrate, that accumulates in tubers amyloplasts. Glucose molecule, produced by photosynthesis, reacts with fructose and forms sucrose, which is the main transport type of carbohydrates in the plant. In the tuber, sucrose is delivered via phloem (apoplast), where it splits into glucose and fructose, which then go to the parenchyma cells. Glucose is a further substrate for the starch synthesis in amyloplasts. However, low temperatures influence on potato tubers leads to starch break down to reducing sugars. In parallel to this process there is happens resynthesis of sucrose to glucose and fructose by acid vacuolar invertase enzyme (β-fructofuranosidase) encoded by Pain-1 gene. Together, these processes lead to an excessive accumulation of monosaccharides in potato tubers. This process also called as cold-induced sweetening. It creates conditions for the intensive formation of melanoidins, which cause a potato tubers darkening, which considerably impairs the commercial quality of the product. Thus, the study Pain-1 gene that encodes the vacuolar invertase (its identification and structure analysis) is an important task required for the search of donors resistant to cold-induced sweetening. The primary task for this is the design and testing of primer combinations that allow to amplify the full-length gene in wild potato species, varieties and lines of cultivated potato. In this work, we develop and test universal primers, that can amplify both full-length orthologs and fragments of the Pain-1 gene, and also select the optimal conditions for carrying out the PCR reaction. Summary. The purpose of research is design and testing of universal primers for PCR amplification of full-length-fructofuranozidase orthologs genes (acid vacuolar invertase) in wild species and potato (Solanum tuberosum) varieties. Starch is the main source of energy and a reserve carbohydrate, that accumulates in tubers amyloplasts. Glucose molecule, produced by photosynthesis, reacts with fructose and forms sucrose, which is the main transport type of carbohydrates in the plant. In the tuber, sucrose is delivered via phloem (apoplast), where it splits into glucose and fructose, which then go to the parenchyma cells. Glucose is a further substrate for the starch synthesis in amyloplasts. However, low temperatures influence on potato tubers leads to starch break down to reducing sugars. In parallel to this process there is happens resynthesis of sucrose to glucose and fructose by acid vacuolar invertase enzyme (β-fructofuranosidase) encoded by Pain-1 gene. Together, these processes lead to an excessive accumulation of monosaccharides in potato tubers. This process also called as cold-induced sweetening. It creates conditions for the intensive formation of melanoidins, which cause a potato tubers darkening, which considerably impairs the commercial quality of the product. Thus, the study Pain-1 gene that encodes the vacuolar invertase (its identification and structure analysis) is an important task required for the search of donors resistant to cold-induced sweetening. The primary task for this is the design and testing of primer combinations that allow to amplify the full-length gene in wild potato species, varieties and lines of cultivated potato. In this work, we develop and test universal primers, that can amplify both full-length orthologs and fragments of the Pain-1 gene, and also select the optimal conditions for carrying out the PCR reaction. In total 6 primer combinations were designed (PainF - PainR, PainF - Pain1exR, Pain1exF - Pain3exR, Pain2inF - Pain2inR, Pain3exF - Pain5exR, Pain5exF - PainR), where PainF - PainR primer combination allowed to amplify a full-sized gene, the rest are internal and will be used in the further fragments sequencing of the β-fructofuranosidase gene. These primers were successfully tested on 15 samples, including five wild species of potato (S. gourlay, S. chacoense, S. pinnatissectum, S. stoloniferum, S. vernei) and ten varieties of Russian and foreign breeding (Gala, Lasunok, Red Scarlet , Rasset Burbank, Miras, Bashkirsky, Zhukovsky ranniy, Matushka, Elizaveta, Sudaryna).

Chromosome pairing and potential for intergeneric recombination in some hypotetraploid somatic hybrids of Lycopersicon esculentum (+) Solanum tuberosum

Genome ◽  
1993 ◽  
Vol 36 (6) ◽  
pp. 1032-1041 ◽  
Author(s):  
J. H. de Jong ◽  
A. M. A. Wolters ◽  
J. M. Kok ◽  
H. Verhaar ◽  
J. van Eden
Keyword(s):  
Solanum Tuberosum ◽  
Lycopersicon Esculentum ◽  
Synaptonemal Complex ◽  
Chromosome Pairing ◽  
Somatic Hybrids ◽  
Cytogenetic Study ◽  
Unreduced Gametes ◽  
Root Tip ◽  
Prophase I ◽  
Metaphase I

Three somatic hybrids resulting from protoplast fusions of a diploid kanamycin-resistant line of tomato (Lycopersicon esculentum) and a dihaploid hygromycin-resistant transformant of a monohaploid potato (Solanum tuberosum) line were used for a cytogenetic study on chromosome pairing and meiotic recombination. Chromosome counts in root-tip meristem cells revealed two hypotetraploids with chromosome complements of 2n = 46 and one with 2n = 47. Electron microscope analyses of synaptonemal complex spreads of hypotonically burst protoplasts at mid prophase I showed abundant exchanges of pairing partners in multivalents involving as many as eight chromosomes. In the cells at late pachytene recombination nodules were found in multivalents on both sides of pairing partner exchanges, indicating recombination at both homologous and homoeologous sites. Light microscope observations of pollen mother cells at late diakinesis and metaphase I also revealed multivalents, though their occurrence in low frequencies betrays the reduction of multivalent number and complexity. Precocious separation of half bivalents at metaphase I and lagging of univalents at anaphase I were observed frequently. Bridges, which may result from an apparent inversion loop found in the synaptonemal complexes of a mid prophase I nucleus, were also quite common at anaphase I, though the expected accompanying fragments could be detected in only a few cells. Most striking were the high frequencies of first division restitution in preparations at metaphase II/anaphase II, giving rise to unreduced gametes. In spite of the expected high numbers of balanced haploid and diploid gametes, male fertility, as revealed by pollen staining, was found to be negligible.Key words: synaptonemal complex, recombination, chromosome pairing, somatic hybrid, Lycopersicon esculentum (+) Solanum tuberosum.

Interspecific somatic hybrids between dihaploidSolanum tuberosumL. and the wild species,S. pinnatisectumDun

1994 ◽  
Vol 45 (10) ◽  
pp. 1433-1440 ◽  
Author(s):  
A.C. Ward ◽  
J.St-J Phelpstead ◽  
A.E. Gleadle ◽  
N.W. Blackhall ◽  
S. Cooper-Bland ◽  
...  
Keyword(s):  
Wild Species ◽  
Somatic Hybrids ◽  
Interspecific Somatic Hybrids

Use of RAPD markers to screen somatic hybrids between Solanum tuberosum and S. brevidens

1993 ◽  
Vol 12 (2) ◽  
Author(s):  
Yong-sheng Xu ◽  
MelodyS. Clark ◽  
Eija Pehu
Keyword(s):  
Solanum Tuberosum ◽  
Rapd Markers ◽  
Somatic Hybrids

Molecular, cytogenetic and morphological characterization of somatic hybrids of dihaploid Solanum tuberosum and diploid S. brevidens

1989 ◽  
Vol 78 (5) ◽  
pp. 696-704 ◽  
Author(s):  
E. Pehu ◽  
A. Karp ◽  
K. Moore ◽  
S. Steele ◽  
R. Dunckley ◽  
...  
Keyword(s):  
Solanum Tuberosum ◽  
Somatic Hybrids ◽  
Morphological Characterization ◽  
Molecular Cytogenetic
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