~ 566 ~ Journal of Pharmacognosy and Phytochemistry 2019; SP1: 566-573 E-ISSN: 2278-4136 P-ISSN: 2349-8234 JPP 2019; SP1: 566-573 Satendra Chaudhary Department of Vegetable Science and Floriculture, CSK Himachal Pradesh Agricultural University, Palampur, Himachal Pradesh, India Viveka Katoch Department of Vegetable Science and Floriculture, CSK Himachal Pradesh Agricultural University, Palampur, (H.P.), India Rakesh Kumar Kapila Department of Vegetable Science and Floriculture, CSK Himachal Pradesh Agricultural University, Palampur, Himachal Pradesh, India Akhilesh Sharma Department of Vegetable Science and Floriculture, CSK Himachal Pradesh Agricultural University, Palampur, Himachal Pradesh, India Shweta Sharma Department of Vegetable Science and Floriculture, CSK Himachal Pradesh Agricultural University, Palampur, Himachal Pradesh, India Correspondence Viveka Katoch Department of Vegetable Science and Floriculture, CSK Himachal Pradesh Agricultural University, Palampur, Himachal Pradesh, India (Special Issue- 1) 2 nd International Conference “Food Security, Nutrition and Sustainable Agriculture - Emerging Technologies” (February 14-16, 2019) Response of Sucrose, PGRs and pretreatments on another culture efficiency in broccoli (Brassica oleracea L. var. italica Plenck.) Satendra Chaudhary, Viveka Katoch, Rakesh Kumar Kapila, Akhilesh Sharma and Shweta Sharma Abstract The present investigation was undertaken to assess the effect of different plant growth regulators (2,4-D and NAA), sucrose concentrations (10, 12, 14 and 16 %) on callus induction frequency and different pretreatment temperatures (cold and heat shock) viz., 0-4°C, 25°C, 27.5°C, 30°C, 32.5°C, 35°C and 37.5°C along with three different incubation periods viz., 24, 48 and 72 hours on Anther culture efficiency in four hybrids of broccoli namely, BR-60, Kendy, Lucky and Fiesta. The highest callus induction frequency was recorded in BR-60 in the treatment T5 (B5 + 0.10 mg/l NAA and 0.50 mg/l 2,4- D + 10 % sucrose). The best response of callus induction frequency was obtained in hybrid BR-60 at 12 per cent concentration of sucrose. The highest anther culture efficiency was observed in BR-60 when anthers were subjected to 35°C for 48 hours using best hormonal combination (T5) and sucrose (12 per cent). The highest calli regeneration frequency was obtained in hybrid Kendy when calli were transferred to HM5 combination (B5 + 0.25 mg/l IAA + 1.0 mg/l BAP + 2.0 mg/l Kin + 3 % sucrose). The rhizogenesis was induced in all the hybrids of broccoli when their calli derived meristemoids were sub- cultured on rooting medium (B5 + 0.5 mg/l NAA + 3 % sucrose). The anther culture response was recorded on the basis of meristemoids. The highest meristemoids were recorded in hybrid Kendy followed by BR-60. Keywords: Broccoli, Anther culture, plant growth regulators, sucrose, pre-treatment Introduction Broccoli (Brassica oleracea L. var. italica Plenck) a member of family Brassicacea is one of the nutritious cole crop. It is known for its taste, flavor, nutritive and medicinal properties. Broccoli contains the compound namely, glucoraphanin, which can be processed into an anti- cancerouus compound sulphoraphane (Kumari and Katoch 2016) [10] . Being a native of Mediterranean region, the diversity of broccoli in India is very less. One of the major bottlenecks for broccoli breeders is the availability of germplasm. Broccoli is cross-pollinated crop and to develop a new inbred through conventional breeding usually takes at least five generations of inbreeding and backcrossing to reach the same goal of recovering genetically stable lines through conventional breeding approaches. For the development of hybrid(s), inbred lines are required and the development of inbred lines is a time consuming task. Androgenesis via anther/microspore culture offers an alternative for the development of new inbred lines in comparatively short-time thereby, reducing the cost incurred in the conventional breeding. The successful proportion of androgenic plantlets production depend on some critical factors including genotype, physiology of the donor plant, microspores/pollen grains stage, culture medium, sucrose concentrations, plant growth regulators and pretreatment temperatures. The present study intends to investigate the effects of different plant growth regulators, different concentrations of sucrose content and various pretreatment temperatures and incubation periods on Anther culture efficiency of broccoli.