Plant regeneration from cell suspension culture in Saccharum officinarum L. and ascertaining of genetic fidelity through RAPD and ISSR markers

Abstract

The aim of this study was to produce sugarcane plantlets from cell suspension culture and study its genetic fidelity using molecular markers. The study was carried out using sugarcane varieties Co 86032 and Q117. Callus cultures of both the varieties were optimized using six different callus induction media. After screening the growth response of callus on six different callus induction media, it was observed that medium no. VI supplemented with 500 mg l−1 of each PVP, Casein hydrolysate and MES buffer showed high amounts of callus in Co 86032 (79.66 ± 0.44%) and Q117 (82.83 ± 1.69%). Addition of PEG 8000 at 2.5% to this medium had a profound impact on inducing somatic embryogenesis in Co 86032 (54.66 ± 1.76%) and Q117 (66.66 ± 2.60%) as compare to control (24.33 ± 1.76%) and (27.33 ± 2.73%), respectively. Cell suspension cultures were established by culturing embryogenic calli in liquid medium showed well established suspension cultures with fever cell aggregates. There was negligible cell division during initial 2 days of incubation and cell count increased rapidly between 2 and 8 days. Further incubation beyond 8 days resulted in a decrease in cell viability. Enhanced callus proliferation in Q117 while enhanced shoot regeneration in Co 86032 was observed from cell suspension culture. The clonal fidelity of in vitro regenerated plants was assessed by using RAPD and ISSR markers. Analysis of the ten RAPD markers indicated that 90.48 and 86.95% true-to-type regenerated plantlets in Co 86032 and Q117, respectively. However, in the ISSR markers, Co 86032 did not show any polymorphism and in the Q117, 92.18% true-to-type plantlets were found. These results confirmed that somaclonal variation occurs during the process of indirect organogenesis and RAPD and ISSR marker based molecular analysis is a suitable method for an early detection of variation in sugarcane.Electronic supplementary materialThe online version of this article (doi:10.1007/s13205-016-0579-3) contains supplementary material, which is available to authorized users.

ANALYSIS OF GENETIC STABILITY OF MICROPROPAGATED SUGARCANE IN DIFFERENT SUBCULTURE FREQUENCIES USING SSR MARKER / Analisis Kestabilan Genetik Tebu Hasil Mikropropagasi pada Beberapa Frekuensi Subkultur Menggunakan Marka SSR

“…The results of the cluster analysis showed that the mother plant samples (control) and the effects of in vitro propagation in one variety had a high level of genetic similarity, which was more than 0.94 except for the PS 862 genotype (Figure 1). The similarity coefficient found in the current study was 0.94 higher than that previously reported by Thorat et al (2017), which analyzed two varieties of sugarcane that produced a genetic similarity coefficient of 0.90 and 0.87, respectively. Also higher than the study of Tiwari et al (2011), which detected a genetic similarity coefficient of more than 85% based on molecular tests using SSR markers.…”

The effects of genotypes and media composition on callogenesis, regeneration and cell suspension culture of chamomile (Matricaria chamomilla L.)

“…Chamomile is known to exist in more than 120 constituents ( McKay & Blumberg, 2006 ), imparting anticancer ( Shoeb et al, 2001a ), analgesic, anti-convulsive, carminative, anti-seizure ( Ghanavatifard, Mohtadi & Masoumiasl, 2018 ), anti-inflammatory, antibacterial, sedative, immune booster, anti-allergic, anti-inflammatory, antiseptic and smooth muscle relaxant properties ( Sotiropoulou, Megremi & Tarantilis, 2020 ). Increased use of herbal medicines has threatened the survival of a large number of medicinally important plant species ( Abbas et al, 2018 ; Mumtaz et al, 2017 ; Thorat et al, 2017 ). However, development of sustainable crop production technologies as well as conservation strategies is inevitable ( Abbas et al, 2020 ; Tahir ul Qamar & Khan, 2017 ).…”

Establishment of a somatic embryo regeneration system and expression analysis of somatic embryogenesis-related genes in Chinese chestnut (Castanea mollissima Blume)

“…However, it did not mean that any mutation of the somatic embryo regeneration system in Chinese chestnut did not occur. Many studies indicated that somaclonal variation occurred during plant regeneration, such as coffee and sugarcane (Landey et al 2015;Thorat et al 2017). No variations were observed during somatic embryogenesis and regeneration in Chinese chestnut, which may be due to a very limited population detected or that the loci identified of genetic fidelity were not sufficient, as only 24 pairs of SSR primers and two gene fragment sequences were identified.…”