Androgenic Responses of Different Pepper (Capsicum annuum L.) Cultivars
Nurgül ERCAN F. AYAR ŞENSOY
Department of Horticulture, Faculty of Agriculture, Akdeniz University 07059 Antalya-TURKEY
*Sorumlu Yazar Geliş Tarihi : 29 Mayıs 2011
nercan@akdeniz.edu.tr Kabul Tarihi : 03 Ağustos 2011 Abstract
Factors such as genotype, donor plant, pre-treatments of buds or anthers, microspore development stage and incubation conditions affect androgenetic response. Eleven different pepper types (4 longy, 1 paprika, 2 bell pepper, 2 capya, 2 long green) were used in this study with the aim of reveal the genotypic response to anther culture. Flower buds excised when the corolla was of the same length as the calyx or slightly longer. Flowers were kept in cold (+ 4oC) and in dark for 24 h. Murashige-Skoog medium with 8 g/l agar and 30g/l sucrose was used. Incubation conditions
were 35 oC for 8 days then transferred to 25 oC with 16/8 h dark-light period with 3000 lux light intensity. Results revealed that pepper types had
different androgenetic responses. A total of 44 embryos were derived from 2398 in vitro cultured anthers of 11 genotypes and 12 plantlets were obtained. Yalova Carliston (long green type) and Kandil (bell type) did not produce any embryos. Sera Demre 8 (longy type) and Odesa (bell type) gave the better androgenic response compare to other pepper genotypes.
Key Words: Anther culture, Capsicum annuum L, genotype, androgenic response
INTRODUCTION
Anther culture has widely been used by breeders as an alternative to the numerous cycles of inbreeding or ‘backcrossing’ usually needed to obtain pure lines [2]. Anther culture is main method of obtaining haploid plants in pepper. The frequency of obtained androgenic plants depends on various factors such as genotype which considerably influence the effectiveness of haploidy; therefore the low rate of haploid plantlets limits the utility of anther culture in pepper breeding.
Mityko and Fari [7] indicated that a decreasing androgenic response was obtained in wax-type, dark green blocky-type, tomato shaped and Cayenne/chilli-type peppers (from 76 to 0 plants/100 cultured anthers). They also reported that although bell peppers generated a high and reproducible response, spice peppers proved to be less or non-responsive.
Nowaczyk and Kisiala [8] reported that the number of plants regenerated from embryos obtained in anther cultures might be increased through optimization of the conditions of their growth.
The most important factor having an effect on the success of the in vitro anther culture is the genotype used. This present study was, therefore, conducted to determine the androgenic response of some pepper cultivar including some Turkish genotypes.
MATERIALS AND METHODS
Pepper cultivars including four longy types (Kekova F1, Demre, Demre sivrisi, Sera- Demre 8), two bell (Kandil, Odesa F1), two capya (Atris, Yağlık biber), one paprika types (DRH-7118 ) and two long green types (Sirena F1, Yalova Carliston) were grown in the greenhouse and used as anther donor genotypes. Flower buds with yellowish-green colored anther were collected when corolla was of the same length as the calyx or somewhat longer [3], and exposed to low temperature pretreatment (4oC and in dark) for 24 hours [1]. Flower buds
were surface sterilized in 10 % sodium hypo chloride solution added with 1-2 drop Tween-20 for 15 minutes. The sterilized buds were rinsed three times with sterile distilled water for five minutes. Anthers of a flower per petri dishes (6 cm in diameter) were excised and laid onto a nutritive medium of Murashige and Skoog (MS) [5] with 4 mg/l NAA and 1 mg/l BA. After adding 30g/l sucrose and 8 g/l agar to MS medium, pH was adjusted to 5.7-5.8. The petri dishes were incubated at 35 oC for 8 days in
the dark [4,6] and then placed at 25 oC with a 16/8 h dark-light
period with 3000 lux-light intensity. Well developed embryos were transferred to hormone free MS medium with 30g/l sucrose and 8 g/l agar for further development. The number and percentage of embryo, callus and plantlet were recorded. Biyoloji Bilimleri Araştırma Dergisi 4 (2): 59-61, 2011
N. Ercan ve F. Ayar Şensoy / Bibad, 4 (2): 59-61, 2011
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RESULTS AND DISCUSSION
Direct embryogenesis was observed in all genotypes investigated in this study. Long green type cv. Yalova carliston and bell type pepper cv. Kandil did not produce any embryos while remaining cultivars produced embryos with different ratio according to genotypes (see table 1). The best results for embryo formation frequency were obtained from the Demre Sivrisi with 7.69% and followed by Odesa F1 with 3.01%. Atris and Sirena cultivars gave the lowest androgenic response with 0.33% and 0.35% respectively. Six embryos of out of 21 embryos in Demre sivrisi were converted to plantlet (Figure 1). Callus formation frequency in all genotypes changed from 17.31% to 44.15%. But any embryo formations from these calluses were
not observed. Callus formed roots or showed abnormality did not continue further development, and gradually turned brown (Figure 2).
The results of this study which is found Kandil pepper cultivar gave no androgenic response are in agreement with Sayılır and Özzambak [10] who reported that cv. Kandil did not produce any embryo. But the other bell type pepper (Odesa F1) used in this study produced embryo and plantlet with the percentage of 3.01 and 22.2, respectively. Studying androgenic response of four breeding lines, seven cultivars and four F1 hybrids Mityko et al. [6], reported that Feherözön, bell pepper variety, (75.8%) and Szechuan 90716 (21.0%) gave the better results compare to the standard cultivar.
Genotypes No of anthercultured No of embryos obtained Embryo formation frequency (%) No of anther formed callus Callus formation frequency(%) No of plantlet obtained from embryos Plantlet formation frequency (%) Atris 312 4 0.33 122 39.10 1 25 Odesa 299 9 3.01 132 44.15 2 22.22 Demre 154 2 1.30 30 19.48 0 0 DRH-7118 275 2 0.73 58 21.10 1 50 Yağlık biber 254 5 1.97 101 39.76 1 20 Yalova carliston 283 0 0.00 49 17.31 0 0 Kandil 262 0 0.00 90 34.35 0 0 Demre sivrisi 273 21 7.69 53 19.41 6 28.57 Sirena 286 1 0.35 84 29.37 1 100 Kekova 467 7 1.50 158 33.83 2 28.57 Sera-Demre 8 442 10 2.26 126 28.51 5 50
Figure 1. a,b,c and d; Direct embryogenesis of peper, f, g, and h; plantlets regenarete from embryos Table 1. Number of cultured anthers, obtained embryos and frequency of responding anthers of 11 genotypes.
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N. Ercan ve F. Ayar Şensoy / Bibad, 4 (2): 59-61, 2011
Figure 2. a; Callus development, b and c; root formation and abnormal development from callus The results of this study which is found different response
between cultivars are in agreement with those Qin and Rotino [9], who reported that anther donor genotype, had a strong effect on the androgenetic response in pepper.
In this study, longy type pepper cultivars Demre Sivrisi and Sera Demre 8 gave the sufficient results to continue future works. These two cultivars are of great importance in Turkey greenhouse pepper growing as currently various breeding programs are being conducted in Demre pepper to improve its agronomic characteristics. Since haploidy studies are promising in Demre pepper, additional research needed to improve embryo formation and plantlet transformation will accelerate the breeding studies conducted. Further studies are also needed in terms of optimization of the nutritive medium, pretreatment and incubation conditions not only for Demre Sivrisi but also per cultivars.
Acknowledgements
This research was supported by Scientific Research Projects Administration Unit of Akdeniz University.
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