Data on optimization of microprojectile bombardment parameters in development of salinity tolerant transgenic lines

This data deals with the optimization of microprojectile bombardment particles for efficient genetic transformation in an indica rice involving AmSOD gene for development of salinity tolerant transgenic lines [1]. In this study, various parameters such as effect of genotypes, helium pressure, osmoticum, explants, flight distance, particle size, particle volume, vacuum, carrier DNA and stopping screen properties have been evaluated to determine their role in transformation of indica rice involving AmSOD gene for development of salinity tolerant Pusa Basmati 1 rice variety. To perform the transformation process, plasmid vector pCAMBIA 1305.2 was used, which harbours GUS Plus™ gene, intron from the castor bean catalase gene, pBR322 ori, kanamycin resistant gene and Xho I site. The transformants have been confirmed using slot blot, polymerase chain reaction and Southern hybridization techniques.

been confirmed using slot blot, polymerase chain reaction and Southern hybridization techniques.
© 2020 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4.0/). Specification Table   Subject Section:1 Agricultural and Biological Sciences Sub section: 06, Agriculture Specific subject area Agricultural Biotechnology intervention made through optimization of microprojectile bombardment particles for efficient transformation in indica rice variety for enhanced tolerance to salinity. Type of data Table  Figure How data were acquired Data were required using the Microprojectile instrument PDS-1000/He system [Bio-Rad, USA (Sanford, 1993)] Based on in vitro culture response only one suitable variety Pusa Basmati 1 among three was selected and subjected to transgenic development involving AmSOD gene. Data were recorded to detect the optimized condition for maximum transient gene expression involving pCAMBIA1305.2 in respect of diverse microprojectile bombardment parameters (physical, chemical and biological). Transient gene expression data through GUS histochemical assay was used and the data were analyzed through statistical software (Microsoft Excel) [2,3]. Data format Analyzed Parameters for data collection In assessing the in vitro culture response to select the most suitable variety with prolific plantlet regeneration, data like callus induction (%), callus proliferation (w/w%), callus health (1e9 scale), shootlet regeneration (%), shootlet health (1e3 scale), root induction (%), hardening (%) and total plantlets grown to maturity were recorded. Similarly, in assessing the role of diverse physical, chemical and biological factors in influencing the transient GUS expression was assessed through GUS histochemical assay [4] involving bombarded calli with binary vector pCAMBIA1305.2 to detect the appropriate condition for microprojectile bombardment involving economically important gene AmSOD [5] in Pusa Basmati 1, especially at the interface of hypervariable climate. Description of data collection Calli were induced by culturing axenic seeds on MS medium [6] supplemented with suitable hormones, which were excised and proliferated on callus proliferation medium containing half dose of 2mg/l 2,4-D as growth regulator was used in the callus induction medium to induce fast growing embryogenic calli. Here in detecting the in vitro culture response to select the most suitable variety with prolific plantlet regeneration, callus induction (%), callus proliferation (w/w%), callus health (1e9 scale), shootlet regeneration (%), shootlet health (1e3 scale), root induction (%), hardening (%) and total plantlets grown to maturity were recorded. Similarly, in assessing the effect of diverse physical, chemical and biological factors in influencing the transient GUS expression was assessed through GUS histochemical assay [4]

Data
Data pertinent to influential parameters such as effect of genotypes, helium pressure, osmoticum, explants, flight distance, particle size, particle volume, vacuum, carrier DNA and stopping screen properties have been evaluated [1] to determine their role in transformation of indica rice involving AmSOD gene for development of salinity tolerant Pusa Basmati 1 rice variety (Table 1).

Effect of genotypes
To determine the effect of genotypes on transformation efficiency three genotypes have been evaluated, of which Pusa Basmati 1 was selected for further experiments (Fig. 1).

Effect of helium pressure
Effect of helium pressure was determined using PDS He 2000 microprojectile instrument (Bio-Rad, USA). Five pressure levels ie., 640e1550 psi have been used in bombardment of 100 randomly selected calli. After completion of 48 h the transient expression of introgressed gene has been determined by GUS expression (Fig. 2).

Effect of osmoticum
To reduce cell viability and sometime necrosis, osmoticum like manitol (Sigma-Aldrich Cat No. M4125) was used to increase the osmolarity. 0.4 M manitol supplemented in the bombardment

Value of the Data
The datasets refers to optimization of microprojectile based particle bombardment in development of tolerant transgenic lines to salinity involving an AmSOD gene from heterologous mangrove plant source into the most important staple food crop rice. This study has determined the role of transformation parameters and corroborated with in vitro culture response, transient gene expression, molecular analyses involving slot, Southern and PCR as well as physiological parameters, hydroponics tests, respectively. Optimized parameters can be used for development of transgenic salt tolerant rice lines with introgression of salt tolerant genes such as p5cs, salT, DREB, HVA1, osmotin etc.  Fig. 3). Dual osmoticum treatments with manitol and sorbitol (0.2 M) were also studied for pre-incubation as shown in (Table 1and Fig . 4).

Effect of explants
To identify most suitable one, different explants were bombarded and GUS expression was recorded under stereo zoom microscope (Nikon, Japan) ( Table 1). Secondary calli was used as suitable explants for microprojectile bombardment for transgenic development (Fig. 5). Microcarrier velocity is intimately related to air resistance that governs transgene delivery and its expression in the recipient system. In the present experiment three flight distances viz. 6, 9, and 12 cm were used to bombard primary calli of 3.0e6.0 cm size with average of 4.5 cm (Table 1 and Fig. 6).   (Table 1 and Fig. 7).

Effect of carrier DNA
To safe guard the intruded DNA from endogenous nuclease inside the recipient calli, Salmon Sperm DNA, Calf thymus DNA, Herring sperm DNA were used as coating material of the microcarriers along with plasmid DNA (pCAMBIA 1305.2 ref.website) in 1:1 ratio. Primary calli of 2.0e3.0 mm size with average of 2.5 mm 2 diameters were bombarded (Fig. 9) (Table 1).