scholarly journals Genetic Stability of the Endangered Species Salix lapponum L. Regenerated In Vitro during the Reintroduction Process

Biology ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 378
Author(s):  
Marzena Parzymies ◽  
Magdalena Pogorzelec ◽  
Katarzyna Głębocka ◽  
Elwira Śliwińska
Keyword(s):  
Genetic Stability ◽  
Inter Simple Sequence Repeat ◽  
Multiplication Rate ◽  
Natural Habitats ◽  
Relict Species ◽  
Flow Cytometric ◽  
Regenerated Plants ◽  
Simple Sequence ◽  
Salix Lapponum

Salix lapponum L. is a boreal relict species, threatened with extinction in Poland. An 80% decrease in the number of its stands was confirmed in the last half-century, so that to prevent the loss of downy willow, attempts were made to reintroduce this species in natural habitats. Micropropagation was chosen as a first stage of its active conservation. S. lapponum shoots were collected and disinfected with NaOCl, AgNO3, or HgCl2 or with a two-step disinfection with NaOCl and then placed on MS medium with BA 1 mg·dm−3 and IBA 0.1 mg·dm−3. Regenerated shoots were cultivated with addition of BA, KIN, or 2iP, alone or in combination with auxins, to find the highest multiplication rate. Inter-simple sequence repeat (ISSR) analysis and flow cytometric analyses were conducted on in vitro regenerated plants to check their genetic stability. Disinfection was quite difficult and the use of HgCl2 was the most efficient. The highest multiplication rate was obtained in presence of KIN at 0.5 mg·dm−3 + IAA at 0.5 mg·dm−3. The analysis confirmed the genome size stability, which is in agreement with the results obtained by ISSR, revealing no somaclonal variation in plantlets and therefore allowing the use of the obtained plants for reintroduction.

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>

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.

Genetic stability and phytochemical profiling of the in vitro regenerated plants of Angelica glauca Edgew.: an endangered medicinal plant of Himalaya

2018 ◽  
Vol 135 (1) ◽  
pp. 111-118 ◽  
Author(s):  
Janhvi Mishra Rawat ◽  
Aakriti Bhandari ◽  
Susmita Mishra ◽  
Balwant Rawat ◽  
Ashok Kumar Dhakad ◽  
...  
Keyword(s):  
Medicinal Plant ◽  
Genetic Stability ◽  
Regenerated Plants ◽  
Endangered Medicinal Plant ◽  
Angelica Glauca

scholarly journals Micropropagation of Inula germanica L. from the Seedlings Explants

2018 ◽  
Vol 46 (1) ◽  
pp. 52-57 ◽  
Author(s):  
Alina TREJGELL ◽  
Monika KAMIŃSKA ◽  
Karolina LISOWSKA ◽  
Andrzej TRETYN
Keyword(s):  
Solid Medium ◽  
Positive Impact ◽  
Shoot Tips ◽  
Fluorescent Light ◽  
First Year ◽  
Naphthaleneacetic Acid ◽  
Multiplication Rate ◽  
Indolebutyric Acid ◽  
Regenerated Plants

This is the first communication of micropropagation system for Inula germanica using seedling explants germinated in vitro. The development of this system gives the possibility of future reintroduction of I. germanica providing a way to stabilize or re-establish its population. Shoot tips and fragments of cotyledons, hypocotyls and roots were isolated from ten-day-old seedlings. Explants were put on MS medium containing 1.0 mg l-1 benzylaminopurine and 0.1 mg l-1 naphthaleneacetic acid and cultured under continuous white fluorescent light (45 μmol.m-2.s-1) at 26 ± 1 °C. The highest percentage of shoot organogenesis (83.3%) was recorded for hypocotyl, while the highest average number of shoots per explant (12.0) was recorded for shoot tips. In subsequent subcultures, multiplication rate decreased to 3.0-4.9 shoots per explant. Less than 19% shoots were able to root on the solid medium without auxins. The highest rooting efficiency (69.3%) was recorded for solid medium supplemented with indolebutyric acid, but growth of roots was inhibited. The percentage of rooted shoots (62.2%) and number of roots per shoot (2.4 per shoot) into the liquid medium were comparable to medium with 0.1 mg·l-1 indolebutyric acid. showing a positive impact on the process of acclimatization. The regenerated plants were able to flowering in the first year after acclimatization. Developed micropropagation system for I. germanica is efficient and can be a useful tool for the active protection of this species.

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.

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