Elite,transformable haploid inducers in maize

The introduction of alleles into commercial crop breeding pipelines is both time consuming and costly.Two technologies that are disrupting traditional breeding processes are doubled haploid(DH)breeding and genome editing(GE).Recently,these techniques were combined into a GE trait delivery system called HI-Edit(Haploid Inducer-Edit).In HI-Edit,the pollen of a haploid inducer line is reprogrammed to deliver GE traits to any variety,obviating recurrent selection.For HI-Edit to operate at scale,an efficient transformable HI line is needed,but most maize varieties are recalcitrant to transformation,and haploid inducers are especially difficult to transform given their aberrant reproductive behaviors.Leveraging marker assisted selection and a three-tiered testing scheme,we report the development of new Iodent and Stiff Stalk maize germplasm that are transformable,have high haploid induction rates,and exhibit a robust,genetically-dominant anthocyanin native trait that may be used for rapid haploid identification.We show that transformation of these elite‘‘HI-Edit”lines is enhanced using the BABYBOOM and WUSCHEL morphogenetic factors.Finally,we evaluate the HI-Edit performance of one of the lines against both Stiff Stalk and non-Stiff Stalk testers.The strategy and results of this study should facilitate the development of commercially scalable HI-Edit systems in diverse crops.

Evaluation of Cobalt Application Combined with Gypsum and Compost as a Regulator of Cabbage Plant Tolerance to Soil Salinity

In response to the global food crisis and the imperative to address soil degradation, the international agricultural policy is actively working to alleviate the adverse impacts of soil salinity. As part of this initiative, a field trial spanning two consecutive seasons (2019/20-2020/21) was conducted under saline conditions. The primary objective was to evaluate the influence of various compost sources, including vermicompost at a rate of 0.5 ton·fed-1 and plant residues compost at a rate of 5.0 ton·fed-1, as main plots. Subplots were established by applying agricultural gypsum, both in the presence and absence of gypsum requirements. Additionally, sub-subplots were created by externally applying cobalt at a rate of 10.0 mg·L-1, with one sub-subplot receiving foliar cobalt application and the other not. The trial sought to assess the growth performance, chemical composition, enzymatic antioxidants, yield, and quality of cabbage plants (Brassica oleracea var. capitata L.) cultivated in saline soil. According to the findings, cabbage plants exhibited the most favorable response in terms of plant height, chlorophyll content, carotene levels, leaf area, nitrogen (N), phosphorus (P), potassium (K), head yield, vitamin C, and total dissolved solids (TDS) when treated with vermicompost, followed by plant compost. Conversely, plants grown without compost exhibited the least improvement in performance. Cabbage treated with agricultural gypsum requirements showed better performance than those without gypsum amendment. Moreover, plants subjected to cobalt spray demonstrated the highest growth, yield, and quality parameters compared to those without cobalt foliar application. In contrast, the control group (plants without the studied treatments) displayed the highest levels of enzymatic antioxidants, specifically catalase and peroxidase. This indicates that soil salinity stress led to an increase in catalase and peroxidase production in cabbage plants as a defense against the harmful impact of reactive oxygen species (ROS) resulting from soil salinity stress. The applied treatments (compost, gypsum, and cobalt) led to a reduction in the cabbage plant’s inherent production of catalase and peroxidase. Generally, the combined treatment of vermicompost × gypsum requirements × cobalt proved effective in mitigating the detrimental effects of soil salinity on cabbage plants. These findings hold significance for farmers and policymakers aiming to enhance agricultural productivity in regions affected by soil salinity. Additionally, further research can explore the long-term effects of these treatments on soil health and crop sustainability.

Correlation and Path Coefficient Analyses in Sugarcane Genotypes of Ethiopia

To study the relations of certain morphological and sugar quality characters with sugarcane yield, an experiment was conducted at Wonji and Metehara Sugar Estates, Ethiopia between March 2012 and October 2013. The experiment, comprising of 400 sugarcane genotypes of which 174 were local that were collected from different regional states of Ethiopia and 226 introduced, was laid out in partial balanced lattice design with two replications. Data was collected on cane yield and its components, sugar yield and sugar quality traits. ANOVA, correlation and path coefficient analysis were done. Analysis of variance revealed significant differences (P < 0.01) for all the characters studied. Cane yield showed strong positive and highly significant (P < 0.01) correlation with millable cane number (r = 0.832), single cane weight (r = 0.528), stalk height (r = 0.517) and sugar yield (r = 0.987). There was also positive significant (P = 0.05) correlation of tiller count and cane diameter with cane yield. Path analysis revealed the highest positive direct effect of millable cane number (0.812) on cane yield followed by single cane weight (0.682) and pol percent (0.550). However, stalk diameter and brix percent had considerable negative direct effects and indirect positive effects through single cane weight on cane yield. Therefore, in view of their significant positive association with cane yield, indirect effects of stalk diameter and brix percent via single cane weight should be considered during selection. Genotypes should be selected on the basis of millable cane number, single cane weight and pol percent for getting higher cane and sugar yield.

iTRAQ-Based Proteomics Investigation of Critical Response Proteins in Embryo and Coleoptile During Rice Anaerobic Germination

Direct-seeding of rice has become popular in recent years due to its low cost and convenience,however,hypoxic condition limits seedling establishment.In this study,weedy rice WR04-6 with high germination ability under anaerobic conditions was used as a gene donor,and we successfully improved the seedling establishment rate of rice cultivar Qishanzhan(QSZ)based on selection of a new rice line R42 from the recombinant inbred line population.R42 inherited high anaerobic germination(AG)ability,and was used for isobaric tags for relative and absolute quantitation(iTRAQ)-based comparative proteomic studies with QSZ to further explore the molecular mechanism of AG.A total of 719 differentially abundant proteins(DAPs)were shared by R42 and QSZ responded to AG,and thus defined as common response DAPs.A total of 300 DAPs that responded to AG were only identified from R42,which were defined as tolerance-specific DAPs.The common response and tolerance-specific DAPs had similar biochemical reaction processes and metabolic pathways in response to anoxic stress,however,they involved different proteins.The tolerance-specific DAPs were involved in amino acid metabolism,starch and sucrose metabolism,tricarboxylic acid cycle pathway,ethylene synthesis pathway,cell wall-associated proteins and activity of active oxygen scavenging enzyme.The in silico protein-protein interactions for the top 60 DAPs indicated that tolerance-specific DAPs had relatively independent protein interaction networks in response to an anoxic environment compared with common response DAPs.The results of physiological indicators showed thatα-amylase and superoxide dismutase activities of R42 were significantly increased under anoxic conditions compared with aerobic conditions.Multiple lines of evidence from western blot,physiological analysis and quantitatDirect seeding of rice has become popular in recent years due to its low cost and convenience,however,hypoxic conditions can limit seedling establishment.In the present study,weedy rice WR04-6 with high germination ability in anaerobic conditions was used as the gene donor and successfully improved the seedling establishment rate of rice cultivar Qishanzhan(QSZ)based on selection of a new rice line R42 from the recombinant inbred line(RIL)population.R42 inherited the had high anaerobic germination(AG)ability,which was used for the isobaric tags for relative and absolute quantitation(iTRAQ)based comparative proteomic studies with QSZ to further explore the molecular mechanism of AG.A total of 719 differentially abundant proteins(DAPs)shared by R42 and QSZ responded to AG and were thus defined as common response DAPs.A total of 300 DAPs that responded to AG were only identified from R42,which were defined as tolerance-specific DAPs.The common response and tolerance-specific DAPs had similar biochemical reaction processes and metabolic pathways in response to anoxic stress,however they involved different proteins.The 300 tolerance-specific DAPs were involved in amino acid metabolism,starch and sucrose metabolism,TCA cycle pathways,ethylene synthesis pathway,cell wall-associated proteins and activity of active oxygen scavenging enzyme.The in silico protein-protein interactions for the top 60 DAPs indicated that tolerance-specific DAPs had relatively independent protein interaction networks in response to an anoxic environment compared with common response DAPs.The results of physiological indicators showed thatα-amylase and SOD activities of R42 were significantly increased under anoxic conditions compared with aerobic conditions.Multiple lines of evidence from western blot,physiological analyses and real-time PCR jointly supported the reliability of proteomics data.In summary,our findings deepened the understanding of the molecular mechanism for the rice response to AG.ive real-time PCR jointly supported the reliability of proteomics data.In summary,our findings deepened the understanding of the molecular mechanism for the rice response to AG.

Genetic Diversity of Quantitative Traits of Sugarcane Genotypes in Ethiopia

Information about the amount and distribution of genetic variation in germplasm collections is important for their efficient management and effective utilization in plant breeding. Therefore this study was conducted to assess genetic diversity of sugarcane germplasm in Ethiopia. An experiment comprising of 400 sugarcane genotypes (174 local and 226 introduced) was conducted between March 2012 and October 2013 at Wonji and Metehara Sugar Estates using partial balanced lattice design with two replications. Data was recorded on 21 quantitative characters which included cane yield and its components, sugar yield and sugar quality traits. ANOVA portrayed highly significant differences (P < 0.01) among the genotypes for 21 quantitative traits. Cluster analysis revealed intra cluster D2 values ranging from 2.16 - 10.60 and inter cluster from 7.24 - 5864. There were six principal components accounting for 79.26% of the total variation in the tested materials. Millable stalk count, single cane weight, stalk diameter, cane yield, sugar yield and sugar quality traits showed high positive loading on the first two PCs and accounted for most of the variation observed among the genotypes. Therefore, this study suggested that the important characters responsible for diversity in the sugarcane genotypes could be grouped in two principal components namely “Yield” and “Quality” with “Yield” traits being comparatively more important than “Quality”. Genotypes clustered for high mean values of various traits could be exploited for further improvement of the crop either through selection or through hybridization. The clusters having high mean value for yield could be selected for yield per se as well.

Heritability and Correlation among Sugarcane (Saccharum spp.) Yield and Some Agronomic and Sugar Quality Traits in Ethiopia

To assess broad sense heritability and phenotypic and genetic correlations among sugarcane yield components, an experiment was conducted at Wonji and Metehara Sugar Estates of Sugar Corporation of Ethiopia during 2012/2013. High broad sense heritability (h2) was detected for stalk diameter (0.730), single cane weight (0.672), millable cane number (0.624), stalk height (0.624) and pol % (0.608), indicating that these traits could be selected for easily. Expected genetic gain of the yield components was moderate to high. All traits had low to high genetic correlations (rg = -0.005 to 0.884) with cane yield and (rg = 0.027 to 0.999) with sugar yield. On average genetic correlations were higher than phenotypic correlations. High Genotypic Coefficient of Variation (GCV), broad sense heritability and expected genetic advance were recorded for stalk diameter, single cane weight and millable cane number. A selection strategy based on these traits could lead to improvement in cane and sugar yield.

植物CRISPR/Cas9多基因编辑载体构建和突变分析的操作方法

CRISPR/Cas9基因组编辑技术是植物基因功能研究与作物改良的有效工具.为此,本实验室开发了高效的CRISPR/Cas9植物多基因编辑载体系统.本载体系统包括6个双元载体和12个含有不同U3/U6启动子的sg RNA中间载体,可满足对单子叶和双子叶植物的遗传转化以及不同抗生素筛选的要求,具有简便、高效,可同时对多基因进行编辑的特点.此外,为了能更高效地应用基因组编辑技术,还开发了一站式在线分析工具包CRISPR-GE.为方便研究人员利用CRISPR/Cas9系统进行植物基因组编辑,本文提供了从靶点选择、CRISPR/Cas9多靶点双元载体构建,以及对靶点突变序列的测序分析等详细的操作方法,以及常见的问题解答.