Influential factors and transcriptome analyses of immature diploid embryo anthocyanin accumulation in maize.

BACKGROUND: Anthocyanins are widely applied as a marker for haploid identification after haploid induction in maize. However, the factors affecting anthocyanin biosynthesis in immature embryos and the genes regulating this process remain unclear. RESULTS: In this study, we analyzed the influence of genetic background of the male and female parents, embryo age and light exposure on anthocyanin accumulation in embryos. The results showed that light exposure was the most crucial factor enhancing the pigmentation of immature embryos. The identification accuracy of haploid embryos reached 96.4% after light exposure, but was only 11.0% following dark treatment. The total anthocyanin content was 7-fold higher in immature embryos cultured for 24 h under light conditions compared to embryos cultured in the dark. Transcriptome analysis revealed that the differentially expressed genes between immature embryos cultured for 24 h in dark and light chambers were significantly enriched in the pathways of flavonoid, flavone, flavonol and anthocyanin biosynthesis. Among the genes involved in anthocyanin biosynthesis, five up-regulated genes were identified: F3H, DFR, ANS, F3'H and the MYB transcription factor-encoding gene C1. The expression patterns of 14 selected genes were confirmed using quantitative reverse transcription-polymerase chain reaction. CONCLUSION: Light is the most important factor facilitating anthocyanin accumulation in immature embryos. After 24 h of exposure to light, the expression levels of the structural genes F3H, DFR, ANS, F3'H and transcription factor gene C1 were significantly up-regulated. This study provides new insight into the factors and key genes regulating anthocyanin biosynthesis in immature embryos, and supports improved efficiency of immature haploid embryo selection during doubled haploid breeding of maize.

Paenibacillus chinensis sp. nov., isolated from maize (Zea mays L.) seeds.

Four Gram-stain positive bacterial strains, designated as 4R1(T), 4R9, 4L13 and 4L18, isolated from seeds of hybrid maize (Zea mays L., Jingke 968), were investigated using a polyphasic taxonomic approach. The cells were found to be facultatively aerobic, motile, spore-forming and rod-shaped. Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolates should be recognised as a species of the genus Paenibacillus, with two close neighbours being Paenibacillus nicotianae YIM h-19(T) (98.41 % similarity) and Paenibacillus hordei RH-N24(T) (98.37 %). The DNA G+C content of strain 4R1(T) was determined to be 51.6 mol %. Its polar lipid profile was found to consist of diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and an unidentified lipid. The predominant respiratory quinone was identified as MK-7 and the major fatty acids were found to be anteiso-C15:0, anteiso-C12:0, anteiso-C13:0 and anteiso-C11:0. Strains 4R1(T), 4R9, 4L13 and 4L18 were clearly distinguished from the reference type strains using phylogenetic analysis, DNA-DNA hybridization and a range of physiological and biochemical characteristics. It is evident from the genotypic and phenotypic data that strains 4R1(T), 4R9, 4L13 and 4L18 represent a novel species of the genus Paenibacillus, for which the name Paenibacillus chinensis sp. nov. is proposed. The type strain is 4R1(T) (=KCTC 33672(T) = CICC 23864(T)).

Paenibacillus zeae sp. nov., isolated from maize (Zea mays L.) seeds.

Four Gram-stain-positive bacterial strains, designated 6R2T, 6R18, 3T2 and 3T10, isolated from seeds of hybrid maize (Zea mays L., Jingke 968) were investigated using a polyphasic taxonomic approach. Cells were aerobic, motile, spore-forming and rod-shaped. Phylogenetic analysis based on 16S rRNA gene sequences indicated that the isolates may represent a novel species of the genus Paenibacillus, the four closest neighbours being Paenibacillus lautus NRRL NRS-666T (97.1 % similarity), Paenibacillus glucanolyticus DSM 5162T (97.0 %), Paenibacillus lactis MB 1871T (97.0 %) and Paenibacillus chibensis JCM 9905T (96.8 %). The DNA G+C content of strain 6R2T was 51.8 mol%. Its polar lipid profile consisted of diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The predominant respiratory quinone was menaquinone 7 (MK-7) and the major fatty acids were anteiso-C15 : 0 and iso-C14 : 0. Strains 6R2T, 6R18, 3T2 and 3T10 were clearly distinguished from the above type strains using phylogenetic analysis, DNA-DNA hybridization, and a range of physiological and biochemical characteristics. It is evident from the genotypic and phenotypic data that strains 6R2T, 6R18, 3T2 and 3T10 represent a novel species of the genus Paenibacillus, for which the name Paenibacillus zeae sp. nov. is proposed. The type strain is 6R2T ( = KCTC 33674T = CICC 23860T).

Effect of Residual Stress Induced by Different Cooling Methods in Heat Treatment on the Fatigue Crack Propagation Behaviour of GH4169 Disc.

In this study, the effects of residual stress induced by three different cooling methods during heat treatment on the crack propagation behaviour of the GH4169 disc were investigated. Different levels of stress fields were induced to the specially designed discs by using air cooling (AC), air jetting cooling (AJC) and water quenching (WQ) methods and were quantitated by numerical simulation. These discs were then subjected to prefabricated cracking, and crack propagation tests were conducted on a spin tester with two load spectrums. Crack growth behaviour was depicted via the surface replica technique and fracture morphology. Regarding the linear superposition of residual stress and centrifugal force, the crack propagation behaviour of different discs was simulated using the FRANC3D software. AJC and WQ introduced compressive residual stress (-259 MPa and -109 MPa, respectively) into the disc compared with the AC method (about -1.5 MPa). The AJC method increases the crack propagation life of the disc by introducing residual compressive stress into the area near the surface of the central hole to inhibit the opening of the crack surface. When the fatigue load was low, this inhibition effect was more significant.

Adjusting sowing date improves the photosynthetic capacity and grain yield by optimizing temperature condition around flowering of summer maize in the North China Plain.

Adjusting the sowing date to optimize temperature conditions is a helpful strategy for mitigating the adverse impact of high temperature on summer maize growth in the North China Plain (NCP). However, the physiological processes of variation in summer maize yield with sowing date-associated changes in temperature conditions around flowering remain to be poorly understood. In this study, field experiments with two maize varieties and three sowing dates (early sowing date, SD1, 21 May; conventional sowing date, SD2, 10 June; delay sowing date, SD3, 30 June) were conducted at Xinxiang of Henan Province in 2019 and 2020. Early sowing markedly decreased the daily mean temperature (T mean), maximum temperature (T max), and minimum temperature (T min) during pre-silking, while delay sowing markedly decreased those temperatures during post-silking. Under these temperature conditions, both varieties under SD1 at 12-leaf stage (V12) and silking stage (R1) while under SD3 at R1 and milking stage (R3) possessed significantly lower malondialdehyde (MDA) content in leaf due to higher activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) compared to SD2. Therefore, SD1 at V12 and R1 stages and SD3 at R1 and R3 stages for both varieties showed significantly higher photosynthetic capacity, including higher SPAD, F v /F m, P n, T r, and G s, which promoted greater pre-silking dry matter (DM) accumulation for SD1 to increase the kernel number, and promoted greater post-silking DM accumulation for SD3 to increase the kernel weight, eventually increased the grain yield of SD1 and SD3 compared to SD2. Results of regression analysis demonstrated that T mean, T max, and T min values from V12 to R1 stages lower than 26.6, 32.5, and 20.3°C are necessary for improving the kernel number, while T mean, T max, T min, and accumulated temperature (AT) values from R1 to R3 stages lower than 23.2, 28.9, 17.3, and 288.6°C are necessary for improving the kernel weight. Overall, optimal temperature conditions around flowering can be obtained by early (21 May) or delay (30 June) sowing to improve the kernel number or kernel weight due to improved photosynthetic capacity, eventually increasing the grain yield of summer maize in the NCP.

[Dynamic characteristics of leaf area index and allocation characters of ecological resources for different yielding spring maize populations].

By using 3-year field experimental results and related meteorological observation data, the dynamic characteristics of leaf area index (LAI) and the allocation characters of ecological resources for different yielding spring maize (Zea Mays L.) population in Huadian of Jilin Province were studied. The results showed that the dynamic characteristics of relative LAI, with the relative growth days of test population, relative effective accumulated temperature, relative sunshine hours and relative rainfall as independent variables, fitted rational formula y = (a + bx) /(1 + cx + dx2), and the regression equation of maize yield with the ratios of growth days before and after silking (x1), effective accumulated temperature before and after silking (x2), rainfall before and after silking (x3), and sunshine hours before and after silking (x4) was y = 5465.19 + 17810.64x(1) - 23236.14x(2) + 4093.41x(3) + 6287.37x(4) (R2 = 0. 8187, P < 0.01), with the effects of these ecological factors on yield being in the sequence of x1 > x2 > x3 > x4 according to the absolute values of partial regression coefficients. In super high yielding (15499.86 kg x hm(-2)) spring maize population, the allocation ratios of x1, x2, x3, and x4 were 1.43, 1.41, 1.44, and 1.40, respectively. Therefore, in Northeast China, appropriate early sowing of spring maize to prolong its growth days with more rainfall and sunshine hours before silking could attain high yielding, and high or super high yield could be achieved when the allocation ratios of x1, x2, x3, and x4 were all about 1.4.