scholarly journals Pengujian Stabilitas Genetik Plantlet Citrumelo Hasil tTCL dari Kultur In Vitro Dengan Menggunakan Teknik Sekuen Berulang

2018 ◽  
Vol 27 (2) ◽  
pp. 165
Author(s):  
Farida Yulianti ◽  
Hidayatul Arisah ◽  
Dita Agisimanto
Keyword(s):  
Tissue Culture ◽  
Genetic Stability ◽  
Simple Sequence Repeat ◽  
Bulk Segregant Analysis ◽  
Cell Layer ◽  
Inter Simple Sequence Repeat ◽  
Transverse Thin Cell Layer ◽  
Simple Sequence

<p>Protokol organogenesis untuk perbanyakan plantlet Citrumelo menggunakan metode transverse thin cell layer (tTCL) batang telah berhasil dikembangkan. Identifikasi stabilitas genetik tanaman hasil kultur jaringan mutlak diperlukan untuk menguji keberadaan off-type. Tujuan penelitian adalah untuk mengetahui potensi primer retrotransposon dan inter simple sequence repeat (ISSR) dalam mendeteksi stabilitas genetik tanaman Citrumelo dari periode kultur yang panjang. Penelitian dilaksanakan pada bulan Juni 2013 sampai dengan Oktober 2015 di Laboratorium Pemuliaan Tanaman, Balai Penelitian Tanaman Jeruk dan Buah Subtropika, Tlekung. Sebanyak empat penanda dengan urutan basa berulang, yaitu retrotransposon dan ISSR digunakan untuk menguji stabilitas genetik plantlet in vitro yang berumur 22 bulan dan untuk mengonfirmasi metode yang dapat diandalkan untuk perbanyakan jeruk Citrumelo yang true-to-type pada masa mendatang. Daun plantlet diseleksi dan diisolasi secara bulk. Amplifikasi dilakukan terhadap DNA dengan sistem bulk segregant analysis (BSA), dan kemudian dipisahkan menggunakan gel agarose. Tanaman in vitro yang sama secara morfologi dapat dibedakan oleh penanda INT-retrotransposon yang mendeteksi adanya kehilangan pita pada grup sampel dengan ukuran 550 bp. Keberadaan retrotransposon dalam genom berlimpah dan aktivasinya diinduksi oleh stres. Kondisi kultur jaringan berpotensi menginduksi aktivasi retrotransposon. Keragaman genetik diperoleh sebesar 2,6%, tetapi masih dapat diterima untuk plantlet yang dihasilkan dari kultur jangka panjang. Plantlet yang digunakan dalam penelitian ini adalah plantlet yang dikulturkan sejak awal tahun 2014 dan telah digunakan untuk mempelajari faktor media dan lingkungan kultur yang efisien pada Citrumelo selama periode 2014–2015. Aktivitas pengkajian variabilitas genetik plantlet yang dihasilkan melalui tTCL batang masih terus dilakukan. Kombinasi protokol dan deteksi berbasis penanda PCR menjadi sarana yang efektif untuk perbanyakan massa benih berkualitas hasil kultur jaringan untuk mendukung progam pemuliaan maupun perbenihan.</p><p>Assessment of genetic stability of long-term cultivation of plantlet derived tTCL Citrumelo using repetitive sequence primers. Regeneration of plantlet from organogenesis of stem transverse thin cell layer (tTCL) was achieved for Citrumelo, a valuable rootstock. Identification of the genetic stability of plant tissue culture is absolutely necessary. The aim of this study was to assess the potential retrotransposons and inter simple sequence repeat (ISSR) primers in detecting the genetic stability of the Citrumelo plantlet derived from tTCL technique. The research was conducted from Juni 2013 until October 2015 in Breeding Laboratory of Indonesian Citrus and Subtropical Fruits Research Institute. A four repetitive based sequences retrotransposon and ISSR marker assays were used to evaluate genetic stability of a group of 22 months old in vitro plantlets and to confirm the most reliable method for true-to-type propagation of Citrumelo. Leaves of plantlets were selected and isolated in bulk. Groups of DNA in bulk segregant analysis (BSA) were amplified and separated using agarose gel. Vitroplants that morphologically similar have been effectively distinguished by a selected primer INT- retrotransposon that detect an deletion band at 550 bp on a line a group of sample. Retrotransposon is abundance through the genome and its activation induced by stress condition. Tissue culture condition was reported potential to induce retrotransposon activation. The genetic variation of 2.6% was acceptable for the culture that produced from long-term. Plantlets used in this study were selected from population induced from early 2014, and employed for studying media as well as environment factors for efficiently organogenesis of citrumelo in period of 2014-2015. However, additional study is on going for evaluating genetic variability from a cycle plantlet production through tTCL of stems. This combination protocol of organogenesis and PCR based markers detection would be powerful tools for mass propagation of high quality seedling derived tissue culture for breeding or cultivation programs.</p>

Caracterización molecular de mutantes de Agave tequilana inducidas con radiación gamma Co60 y su efecto en la acumulación de fructooligosacáridos

2020 ◽  
Vol 30 ◽  
pp. 1-11
Author(s):  
Alejandro Ángeles-Espino ◽  
Hilda Eréndira Dimas-Estrada ◽  
David Ramírez-Alvarado ◽  
José Manuel Cruz-Rubio ◽  
Paola Andrea Palmeros-Suárez ◽  
...  
Keyword(s):  
Simple Sequence Repeat ◽  
Inter Simple Sequence Repeat ◽  
Agave Tequilana ◽  
Sequence Repeat ◽  
Simple Sequence

La variabilidad genética en Agave tequilana es limitada debido a la manera clonal de reproducción en campo. En el presente trabajo se indujeron mutaciones en plantas de Agave tequilana micropropagadas in vitro vía embriogénesis indirecta empleando radiación ionizante con rayos gamma Co60 a las dosis de 10 Gy, 15 Gy, 20 Gy y 25 Gy. Las plántulas regeneradas fueron nuevamente tratadas a 10 Gy y 20 Gy para obtener poblaciones clonales simple y doblemente irradiadas. Se evidenció variabilidad genética de las poblaciones irradiadas mediante marcadores Inter Simple Sequence Repeat (ISSR) agrupándolas según la dosis de radiación. Las mutantes simples y dobles a 20 Gy aumentaron significativamente el contenido de fructosa y sacarosa en tallo, respecto al control sin irradiar, y la síntesis de fructooligosacáridos con grados de polimerización de 3, 4, 5 incrementó, lo que sugiere la inducción de los genes 1-SST y 1-FFT en las variantes de Agave tequilana.

Somatic chimerism, genetic inheritance, and mapping of the fleshless berry (flb) mutation in grapevine (Vitis vinifera L.)

Genome ◽  
2006 ◽  
Vol 49 (7) ◽  
pp. 721-728 ◽  
Author(s):  
L Fernandez ◽  
A Doligez ◽  
G Lopez ◽  
M R Thomas ◽  
A Bouquet ◽  
...  
Keyword(s):  
Vitis Vinifera ◽  
Ssr Markers ◽  
Simple Sequence Repeat ◽  
Bulk Segregant Analysis ◽  
Cell Layer ◽  
Vitis Vinifera L ◽  
Sequence Repeat ◽  
Wild Type ◽  
Target Region ◽  
Simple Sequence

The fleshless berry (flb) mutation of grapevine (Vitis vinifera L. 'Ugni Blanc') impairs the differentiation and division of inner mesocarp cells responsible for flesh in grapevine berries. In order to study the inheritance of the mutation and to map the flb locus, 5 segregating populations were created. Progeny plants were classified as mutant or wild type by scoring for the presence of an ovary phenotype associated with the Flb– phenotype at anthesis. Phenotypic segregation revealed the involvement of a single dominant allele that was heterozygous in the original mutant. Through bulk segregant analysis, microsatellite (simple sequence repeat (SSR)) markers linked to the mutation were identified, and the flb locus was assigned to linkage group 18. The locus position was then refined by analyzing individual progeny and the segregation of SSR markers in the target region with the closest marker 5.6 cM distant from the flb locus. All progeny with the Flb– ovary phenotype differed from the original fleshless berry mutant in that no berries formed after anthesis. Our data suggest that the original mutant plant was a chimera with the mutated allele present in only 1 cell layer (L2 layer) of the ovary and berry.Key words: Vitis vinifera, mutation, genetic mapping, chimerism.

scholarly journals Field Performance and Genetic Stability of Micropropagated Gooseberry Plants (Ribes grossularia L.)

Agronomy ◽  
2020 ◽  
Vol 11 (1) ◽  
pp. 45
Author(s):  
Danuta Wójcik ◽  
Aleksandra Trzewik ◽  
Danuta Kucharska
Keyword(s):  
Genetic Stability ◽  
Fragment Length Polymorphism ◽  
Field Performance ◽  
Inter Simple Sequence Repeat ◽  
Planting Material ◽  
Small Fruit ◽  
Micropropagated Plants ◽  
Growth Vigor ◽  
Simple Sequence

Gooseberry (Ribes grossularia L.) is a small fruit crop producing valuable fruits, which is constantly gaining importance. In vitro propagation of this species can significantly support the production of virus-free planting material and accelerate the introduction of new cultivars to the market. The aim of presented study was to assess field performance and genetic stability of micropropagated plants (MPs) of four gooseberry cultivars, “Captivator”, “Hinnonmaki Rot”, “Invicta”, and “Resika”. The growth vigor and yield of MPs and plants propagated by standard methods from softwood cuttings (ST) were evaluated in a field experiment. Microscopic observations of the number and length of the stomata of MP and ST plants were carried out. Two DNA-based techniques, amplified fragment length polymorphism (AFLP) and inter simple sequence repeat (ISSR), were used to assess genetic stability of MP plants. For analysis of genetic stability of ST plants, the ISSR technique was applied. For three cultivars, Captivator, Hinnonmaki Rot, and Invicta, the plants’ growth vigor and fruit yield were greater in MP plants than in ST plants. In the case of Resika, most of these parameters were higher in ST plants. Microscopic observations of the stomata indicated a lack of differences in the length between MP and ST plants, while the stomata frequency on leaves of MP plants was higher than that of ST plants. The genetic variability of MP plants, assessed by AFLP, ranged from 0.35% for Hinnonmaki Rot to 2.12% for Resika. The results of ISSR analysis of MP plants showed variability from 0% in the case of Hinnonmaki Rot and Resika to 4% and 8.69% for Captivator and Invicta, respectively. No polymorphism was detected among ST plants of all analyzed gooseberry cultivars.

scholarly journals Molecular Characterization of Verbascum anisophyllum (Scrophulariaceae) Genetic Resources Through Inter-Simple Sequence Repeat (ISSR) Markers

2017 ◽  
Vol 12 (2) ◽  
pp. 1934578X1701200 ◽  
Author(s):  
Galya Petrova ◽  
Stefan Petrov ◽  
Svetlana Bancheva
Keyword(s):  
Genetic Diversity ◽  
Genetic Resources ◽  
Simple Sequence Repeat ◽  
Inter Simple Sequence Repeat ◽  
Issr Markers ◽  
Sequence Repeat ◽  
Data Book ◽  
Simple Sequence

Verbascum (Mullein) flowers are highly valued as natural remedy for various respiratory diseases. Verbascum anisophyllum Murb. is a Balkan endemic, protected by law and included in the Bulgarian Red Data Book as “Critically Endangered”. Thus, a strict conservation policy and a reliable evaluation of its genetic resources are required, considering its narrow distribution range and the increasing risk from destruction of its habitats. Here, we used Inter-simple sequence repeat (ISSR) markers to characterize the genetic diversity and to assess the genetic differentiation between the existing populations of Verbascum anisophyllum in Bulgaria. The level of genetic diversity found herein clearly indicates a long-term potential for adaptability of this endangered plant. Our findings provide important knowledge of population genetic structure of this species, thus representing a strategy for its efficient conservation and utilization.

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