Assessment for the Incidence of Number of Subcultures on Genotype Stability for In Vitro Plantlets of Yam (Dioscorea spp.) Using RAPD Markers

Agbidinoukoun, Arnaud; Missihoun, Antoine Abel; Akonde, Pierre; Sagbadja, Hurbert Adoukonou; Agbangla, Clément; Ahanhanzo, Corneille

doi:10.20546/ijcrbp.2017.404.006

Cited by 3 publications

(7 citation statements)

References 22 publications

(20 reference statements)

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“…Variasi somaklonal tidak terjadi pada sample klon jati hasil kultur jaringan yang diujikan. Banyak faktor yang menyebabkan tidak ditemukannya variasi somaklonal pada tanaman hasil kultur jaringan, seperti sumber eksplan, perlakuan zat pengatur tumbuh, lamanya kultur dan sensitivitas penanda DNA yang digunakan (Sarmento, Martins, & Oliveira, 2005;Agbidinoukoun et al, 2017;Sarmah, Sutradhar, & Singh, 2017). Sumber eksplan dari jaringan yang masih aktif membelah seperti kambium kemungkinan dapat mengurangi terjadinya variasi.…”

Section: Bunclassified

“…Frekuensi sub kultur dianggap sebagai salah satu penyebab munculnya variasi somaklonal. Variasi somklonal meningkat dengan semakin banyaknya frekuensi sub kultur (Akdemir et al, 2016); Roostika, Khumaida, & Ardie, 2015;Agbidinoukoun et al, 2017). Variasi terjadi setelah sub kultur ke-4 (Bairu et al, 2011) kultur pisang (Krishna et al, 2016).…”

Section: Keteranganunclassified

“…Jumlah primer RAPD yang digunakan untuk menguji variasi somaklonal klon hasil kultur jaringan berkisar antara 2 (Chinmayee, Aadeshkumar, & Monica, 2012;Hattab, El-Kaaby, & Saadon Abdulhadi Al-Ajeel, 2017 ), puluhan (Roostika et al, 2015) hingga ratusan primer (Sarmento et al, 2005). Selain itu, jumlah sampel yang digunakan tidak terlalu banyak, berkisar antara 1 sampai 4 sampel klon (Darwesh et al, 2017;Roostika et al, 2015;Khoddamzadeh et al, 2010;Agbidinoukoun, Missihoun, Akonde, Sagbadja, Agbangla, & Ahanhanzo, 2017). Penelitian sebelumnya melaporkan bahwa pada region tertentu dalam untai DNA mempunyai titik (hot spot) instabilitas sehingga akan dengan mudah mendeteksi variasi somaklonal meskipun menggunakan primer dan sampel dengan jumlah terbatas (Linacero, Alves, & Vazquez, 2000;Hattab et al, 2017).…”

Section: Keteranganunclassified

“…Sebaliknya, beberapa penelitian melaporkan bahwa variasi somaklonal tidak ditemukan meskipun menggunakan ratusan primer RAPD. Variasi somaklonal tidak ditemukan karena terjadi kestabilan genetik pada target fragment DNA (Agbidinoukoun et al, 2017). Usaha untuk menambah jumlah primer RAPD dilakukan untuk memvalidasi terjadinya kestabilan genetik (Sarmento et al, 2005).…”

Section: Keteranganunclassified

“…Penanda RAPD sering diaplikasikan untuk mengidentifikasi variasi somaklonal (Roostika et al, 2015;Gadakh et al, 2017), meskipun ampilifikasi penanda RAPD sering tidak konsisten karena sekuen primer berukuran pendek (10 bp) dan menempel secara acak pada untai DNA. (Sarmento et al, 2005;Agbidinoukoun, Missihoun, Akonde, Sagbadja, Agbangla, & Ahanhanzo, 2017). Untuk meminimalis kelemahan ini, hal yang perlu dilakukan adalah harus mengacu pada protokol yang baku dan metode pemilihan lokus yang tepat (Bairu et al, 2011).…”

Section: Keteranganunclassified

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PENGUJIAN PENANDA RANDOM AMPLIFIED POLYMORPHISM DNA UNTUK MENGETAHUI KESTABILAN GENETIK KLON JATI (Tectona grandis)

Nurtjahjaningsih¹,

Herawan²,

Rachma

et al. 2018

JPTH

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This study aimed to test RAPD markers to assess genetic stability of teak clones. Two experimental steps were carried out. First, nine RAPD markers were screened to verify the level of polymorphic loci using non clone samples; second, the polymorphic loci were applied to test genetic stability of clones from tissue culture. To test polymorphism levels of the primers, DNA was isolated from eight leaf samples that were collected from a seed orchard located at Watusipat, Gunung Kidul. To verify genetic stability of clones, DNA was isolated from leaf samples of 24 ramets of 3 clones after second sub-culturing. Results showed that amplification of 5 out of 9 RAPD primers were consistent and produced 12 polymorphic loci. The number of polymorphic alleles per locus ranged between 1 and 3; the allele sizes were between 400 and 1050 base pairs (bps). The percentage of polymorphic loci was 100%; it meant that overall loci have high polymorphism level. Based on these loci showed that the 24 ramets are clones; there was no somaclonal variation or high genetic stability. However, these loci need to be validated using more stable DNA markers. ABSTRAKPenelitian ini bertujuan untuk menguji penanda RAPD pada kestabilan genetik klon jati. Pengujian ini melalui dua tahap penelitian yaitu memilih penanda RAPD yang mengamplifikasikan lokus polimorfik pada sampel bukan klon, kemudian target lokus polimorfik tersebut diujikan pada sampel klon hasil kultur jaringan. Sampel DNA bukan klon berasal dari kebun benih jati di Watusipat, Gunung Kidul. Sampel DNA klon berasal dari 24 ramet, dari 3 klon setelah 2 kali sub kultur. Hasil penelitian menunjukkan bahwa 5 dari 9 penanda RAPD pada sampel bukan klon, bersifat konsisten dan menghasilkan 12 lokus polimorfik. Jumlah alel polimorfik per lokus berkisar antara 1 sampai dengan 3; ukuran alel berkisar antara 400 sampai 1050 pasangan basa (bp). Persentase lokus polimorfik bernilai 100% menunjukkan bahwa seluruh lokus mempunyai nilai polymorphism tinggi. Berdasarkan 12 lokus tersebut, 24 ramet menunjukkan genotipe yang sama pada masing-masing klonnya. Namun demikian, target lokus tersebut masih harus divalidasi menggunakan penanda DNA yang sifatnya lebih stabil. Kata kunci: kultur jaringan, pemilihan primer, lokus polimorfik, variasi somaklonal

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Section: Bunclassified

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PENGUJIAN PENANDA RANDOM AMPLIFIED POLYMORPHISM DNA UNTUK MENGETAHUI KESTABILAN GENETIK KLON JATI (Tectona grandis)

Nurtjahjaningsih¹,

Herawan²,

Rachma

et al. 2018

JPTH

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Short Communication: Genetic similarity analysis of in vitro cultivated pineapple (Ananas comosus) from Sipahutar, North Sumatra, Indonesia using ISSR markers

Harahap

NUSYIRWAN²,

AFIVA³

et al. 2022

Biodiversitas

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Abstract. Harahap F, Nusyirwan N, Afiva A, Prasetya E, Suriani C, Hasibuan RFM, Poerwanto R. 2022. Short Communication: Genetic similarity analysis of in vitro cultivated pineapple (Ananas comosus) from Sipahutar, North Sumatra, Indonesia using ISSR markers. Biodiversitas 23: 5623-5628. Sipahutar pineapple from North Sumatra, Indonesia, is considered difficult to breed, resulting in a lack of seed availability. However, tissue culture allows the emergence of somaclonal variations, especially in plantlets undergoing several subculture stages. This study aimed to identify somaclonal variations in Sipahutar pineapple seedlings generated from in vitro culture using Inter Simple Sequence Repeat (ISSR) markers. Fifteen accessions were used in this study consisting of Sipahutar pineapple grown in its original environment, a laboratory environment, acclimatized, and in vitro cultured. Total of 10 primers, of which seven primers detected polymorphisms among the population. This study resulted in a low genetic similarity among the Sipahutar pineapple population. The genetic diversity of Sipahutar pineapple plants propagated by in vitro culture had a similarity level of 75%-94%, showing somaclonal variations in vitro culture. The genetic similarity of pineapple accessions was divided into two main clusters, namely cluster A consisted of 11 accessions and cluster B consisted of 4 accessions. Cluster separation does not correlate to the source of plant materials studied.

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Analysis of genetic diversity in four Sudanese provenances of Balanites aegyptiaca (L.) Del. based on random amplified polymorphic DNA (RAPD) marker

Freigoun¹,

Elagib²,

Raddad³

2020

Afr. J. Biotechnol.

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Balanites aegyptiaca (L.) Del. (Heglig) is a multipurpose tree with considerable potential. The wide range of environments under which it is growing suggests high pattern of variation among and within locations. The scrutiny was undertaken to assess genetic diversity among and within four geographical genotypes of Heglig growing naturally in four different regions of Sudan based on their seed performance using random amplified polymorphic DNA (RAPD) markers. The DNA was extracted from dried leaf materials and subjected to PCR using ten primers. A total of 57 bands were scored and 42 were polymorphic with polymorphism level ranging from 50 to 87.5% and a mean of 71.7%. Cluster analysis using NTSYS-PC software, showed three main clusters. The dissimilarity values ranged between 77 and 93%. Effective gene flow among the three populations of Obied, Damazin and Gedaref and limited gene flow with Genaina was observed. Populations of B. aegyptiaca from different provenances were confirmed to have significant genetic diversity. Results indicate that RAPD could be efficiently used for studying genetic variation of B. aegyptiaca. The study recommends that local provenances of B. aegyptiaca should be properly conserved and immediate efforts to be made to widen the genetic base through research and collection from other regions.

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Assessment for the Incidence of Number of Subcultures on Genotype Stability for In Vitro Plantlets of Yam (Dioscorea spp.) Using RAPD Markers

Cited by 3 publications

References 22 publications

PENGUJIAN PENANDA RANDOM AMPLIFIED POLYMORPHISM DNA UNTUK MENGETAHUI KESTABILAN GENETIK KLON JATI (Tectona grandis)

PENGUJIAN PENANDA RANDOM AMPLIFIED POLYMORPHISM DNA UNTUK MENGETAHUI KESTABILAN GENETIK KLON JATI (Tectona grandis)

Short Communication: Genetic similarity analysis of in vitro cultivated pineapple (Ananas comosus) from Sipahutar, North Sumatra, Indonesia using ISSR markers

Analysis of genetic diversity in four Sudanese provenances of Balanites aegyptiaca (L.) Del. based on random amplified polymorphic DNA (RAPD) marker

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