Quantitative Assessment of the Dominant Genome in Fusant Cultures

Abstract

Selection of fusant cultures, producers of bioactive compounds is of significant importance for the biotechnology industry.Protoplast fusion between prototrophic strains of Aspergillus oryzae PP, producer of alfa-amylase and Apergillus awamori K-1, producer of xylanase was obtained. The interspecific nature of the fusants was determined by morphology and screening for combined enzymatic activity. The aim of this investigation was to quantitate the dominant parental genome in the fusants by applying whole genome typing strategy based on the Amplified Fragment Length Polymorphisms (AFLP) technique. In addition we investigated the genome decomposition of the fusant cultures from first to fourth generation. Our results demonstrate that the appearances of the fusants varied but they basically resembled Aspergillus awamori. On the bases of more than 80 polymorphic markers, the genomic analysis confirmed that Aspergillus awamori is the dominant genome (>85%) in the fusants. The stability of the fusants was examined by successive subcultures. The genomic decomposition of the fusants between first and forth generation is as little as 3%. From our studies we could conclude that AFLP is an efficient and discriminatory method for evaluation of the dominant genome in fusant cultures and the extent of genomic decomposition in the generations.

Karakterisasi Dan Identifikasi Molekuler Fusan Hasil Fusi Protoplas Interspesies Chlorella pyrenoidosa dan Chlorella vulgaris Menggunakan 18SrDNA

“…Fusi protoplas merupakan suatu metode untuk menggabungkan dua atau lebih sel sehingga diperoleh kombinasi sel baru dengan bantuan aliran listrik atau zat kimia (Mariska dan Husni, 2002; (Kusumaningrum dan Zainuri, 2014; Kusumaningrum dan Zainuri 2015a; Kusumaningrum dan Zainuri 2015b; Kusumaningrum dan Zainuri 2015c). Karakter morfologi dari gabungan keduanya dibandingkan dengan induk menjadi faktor yang biasa digunakan oleh peneliti lain (Panaiotov et al, 2009). Namun, karakterisasi secara konvensional ini memiliki kemungkinan bahwa mikroalga yang memiliki fenotip sama teridentifikasi menjadi spesies yang sama, padahal keduanya belum tentu secara genetik memiliki kesamaan.…”

Improvement of Nutrition Production by Protoplast Fusion Techniques in Chlorella vulgaris

“…Application of protoplast fusion process for microalgae Chlorella had conducted to improve their carotenoid for animal aquatic supplement [10]. The use of fusant as a natural feed supplement is potential to overcome synthetic feed [1,[16][17][18][19]. Application of fusant as natural feed supplement on Penaeus monodon post larvae had raised the weight growth, and survival from microbial diseases and immune response in wide range of salinity [1,20].…”