Sorghum has been considered a recalcitrant plant in vitro and suffers from a lack of regeneration protocols that function broadly and efficiently across a range of genotypes. This study was initiated to identify differential genotype-in vitro protocol responses across a range of bioenergy sorghum parental lines and the common grain sorghum genotype Tx430 in order to characterize response profiles for use in future genetic studies. Two different in vitro protocols, LG and WU, were used for comparisons. Distinct genotype-protocol responses were observed, and the WU protocol performed significantly better for plantlet regeneration. Most bioenergy genotypes performed as well, if not better than Tx430, with Rio and PI329311 as the top regenerating lines. Genotypes displayed protocol-dependent, differential phenolic exudation responses, as indicated by medium browning. During the callus induction phase, genotypes prone to medium browning exhibited a response on WU medium which was either equal or greater than on LG medium. Genotype- and protocol-dependent albino plantlet regeneration was also noted, with three of the bioenergy genotypes showing albino plantlet regeneration. Grassl, Rio and Pink Kafir were susceptible to albino plantlet regeneration, with the response strongly associated with the WU protocol. These bioenergy parental genotypes, and their differential responses under two in vitro protocols, provide tools to further explore and assess the role of genetic loci, candidate genes, and allelic variants in the regulation of in vitro responsiveness in sorghum.
25Sorghum has been considered a recalcitrant plant in vitro, and suffers from a lack of 26 regeneration protocols that function broadly and efficiently across a range of genotypes. This study 27 was initiated to identify differential genotype-in vitro protocol responses across a range of 28 bioenergy sorghum bioenergy parental lines, in order to characterize response profiles for use in 29 future genetic studies. Seven bioenergy sorghum genotypes were compared, along with the 30 common grain sorghum genotype Tx430, for their in vitro regeneration responses using two 31 different in vitro protocols, LG and WU. All genotypes displayed some level of response during 32 in vitro culture with both protocols. Distinct genotype-protocol responses were observed, with the 33 WU protocol significantly better for plantlet regeneration. All bioenergy genotypes, with the 34 exception of Chinese Amber, performed as well, if not better than Tx430, with Rio and PI329311 35 the top regenerating lines. Genotypes displayed protocol-dependent, differential phenolic 36 exudation responses, as indicated by medium browning. During the callus induction phase, 37 genotypes prone to medium browning exhibited a response on WU medium which was either equal 38 or greater than on LG medium, with Pink Kafir and PI329311 the most prone to medium browning. 39 Genotype-and protocol-dependent albino plantlet regeneration was also noted, with three of the 40 bioenergy genotypes showing albino plantlet regeneration. Grassl, Rio and Pink Kafir were 41 susceptible to albino plantlet regeneration, with the response strongly associated with the WU 42 protocol. Pink Kafir displayed the highest albino formation, with close to 25% of regenerating 43 explants forming albino plantlets. 44 45 46 47 48 49 50 51 52 53 Sorghum [Sorghum bicolor (L.) Moench] ranks fifth of the major grain crops in production, 54 area harvested, and yield worldwide (FAOSTAT Database 2017), and more than 300 million 55 people use it as a staple food, particularly in developing semiarid tropical regions (Kebede et al.56
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