Increasing protein content and digestibility in sorghum grain with a synthetic biology approach

Despite great genetic diversity, sorghum grain consistently suffers from poor protein digestibility. The physicochemical packaging of protein bodies which consist of protease-resistant β- and γ-kafirin is considered a major obstacle. A synthetic β-kafirin gene, which shares the endosperm-specific promoter and signal peptide with the native β-kafirin gene (Sobic.009G001600.1), was transformed into sorghum inbred line Tx430. The gene was modified with ten additional proteolytic sites. These sites were designed to be amenable to cleavage by pepsin and/or chymotrypsin proteinases. Five independent transgenic lines were regenerated by microprojectile transformation. Notably, considerably more protein was observed in the peripheral endosperm of transgenic lines under scanning electron microscopy. Microscopy revealed invaginated or irregularly shaped protein bodies in the endosperm of transgenic lines. Grains of transgenic lines contained 11–37% more protein, which was 11–21% more pepsin digestible and 7–25% more chymotrypsin digestible than Tx430. Additionally, the abundant synthetic β-kafirin protein (5.6% of total protein) was detected by mass spectrometry data analysis in the transgenic line 9-1. Field-grown homozygous transgenics retained higher protein content, larger seed size and no reduction in grain number per plant. The results illustrated that plant synthetic biology could play an important role in improving sorghum nutritional value

A radiation-hardened SOI-based FPGA

A radiation-hardened SRAM-based field programmable gate array VS1000 is designed and fabricated with a0.5μm partial-depletion silicon-on-insulator logic process at the CETC58th Institute. The new logic cell(LC), with a multi-mode based on3-input look-up-table(LUT), increases logic density about12% compared to a traditional4-input LUT The logic block(LB), consisting of2 LCs, can be used in two functional modes: LUT mode and distributed read access memory mode. The hierarchical routing channel block and switch block can significantly improve the flexibility and routability of the routing resource. The VS1000 uses a CQFP208 package and contains392 reconfigurable LCs,112 reconfigurable user I/Os and IEEE1149.1 compatible with boundary-scan logic for testing and programming. The function test results indicate that the hardware and software cooperate successfully and the VS1000 works correctly. Moreover, the radiation test results indicate that the VS1000 chip has total dose tolerance of100 krad(Si), a dose rate survivability of1.5×1011 rad(Si)/s and a neutron fluence immunity of1×1014 n/cm2.?2011 Chinese Institute of Electronics

Genetic modification of PIN genes induces causal mechanisms of stay-green drought adaptation phenotype

The stay-green trait is recognised as a key drought adaptation mechanism in cereals worldwide. Stay-green sorghum plants exhibit delayed senescence of leaves and stems, leading to prolonged growth, a reduced risk of lodging and higher grain yield under end-of-season drought stress. More than 45 quantitative trait loci (QTL) associated with stay-green have been identified, including two major QTL (Stg1 and Stg2). However, none of the contributing genes that regulate functional stay-green are known. Here we show that the PIN FORMED family of auxin efflux carriers induce some of the causal mechanisms driving the stay-green phenotype in sorghum, with SbPIN4 and SbPIN2 located in Stg1 and Stg2, respectively. We found that 9 of 11 sorghum PIN genes aligned with known stay-green QTL. In transgenic studies, we demonstrated that PIN genes located within the Stg1 (SbPIN4), Stg2 (SbPIN2) and Stg3b (SbPIN1) QTL regions acted pleiotropically to modulate canopy development, root architecture and panicle growth in sorghum, with SbPIN1, SbPIN2 and SbPIN4 differentially expressed in various organs relative to the non stay-green control. The emergent consequence of such modifications in canopy and root architecture is a stay-green phenotype. Crop simulation modelling shows that the SbPIN2 phenotype can increase grain yield under drought

Genetic modification of PIN genes induces causal mechanisms of stay-green drought adaptation phenotype

The stay-green trait is recognised as a key drought adaptation mechanism in cereals worldwide. Stay-green sorghum plants exhibit delayed senescence of leaves and stems, leading to prolonged growth, a reduced risk of lodging and higher grain yield under end-of-season drought stress. More than 45 quantitative trait loci (QTL) associated with stay-green have been identified, including two major QTL (Stg1 and Stg2). However, none of the contributing genes that regulate functional stay-green are known. Here we show that the PIN FORMED family of auxin efflux carriers induce some of the causal mechanisms driving the stay-green phenotype in sorghum, with SbPIN4 and SbPIN2 located in Stg1 and Stg2, respectively. We found that 9 of 11 sorghum PIN genes aligned with known stay-green QTL. In transgenic studies, we demonstrated that PIN genes located within the Stg1 (SbPIN4), Stg2 (SbPIN2) and Stg3b (SbPIN1) QTL regions acted pleiotropically to modulate canopy development, root architecture and panicle growth in sorghum, with SbPIN1, SbPIN2 and SbPIN4 differentially expressed in various organs relative to the non stay-green control. The emergent consequence of such modifications in canopy and root architecture is a stay-green phenotype. Crop simulation modelling shows that the SbPIN2 phenotype can increase grain yield under drought

Additive effects of three auxins and copper on sorghum in vitro root induction

A healthy root system is vital for tissue culture plantlet survival and rapid adaptation from the in vitro microenvironment to glasshouse conditions. Optimization of the root induction medium is an effective way to promote root induction and elongation. Levels of three auxins (alpha-naphthaleneacetic acid [NAA], 3-indoleacetic acid [IAA], and 3-indolebutyric acid [IBA]) and copper sulfate (CuSO4) have been investigated in a series of experiments with a sorghum inbred line, Tx430. Significant improvement in root proliferation and shoot growth were observed on Murashige and Skoog (MS) medium supplemented with 1 mu mol/L CuSO4, 1 mg/L NAA, 1 mg/L IAA, and 1 mg/L IBA. On average, one explant (the original in vitro-derived shoot) of Tx430 regenerated 56.7 roots, which was 20-fold higher on the optimal medium than on MS control medium. Another tested genotype SA281 showed similar response patterns as Tx430 across media. In addition, 100% of Tx430 and SA281 plantlets originating from the optimized root induction medium all survived after being transferred to potting soil in the glasshouse. The results demonstrate that a combination of auxins (NAA, IAA, and IBA) and CuSO4 together at optimal concentrations provide additive effects on promoting root proliferation and explant growth of in vitro sorghum in root induction medium, and subsequently resulted in 100% survival rate of plantlets ex tissue culture. Compared with two published and frequently used root induction media, the optimized medium significantly enhanced root induction and plantlet growth