Improvements of TKC Technology Accelerate Isolation of Transgene-Free CRISPR/Cas9-Edited Rice Plants

Elimination of the CRISPR/Cas9 constructs in edited plants is a prerequisite for assessing genetic stability, conducting phenotypic characterization, and applying for commercialization of the plants. However, removal of the CRISPR/Cas9 transgenes by genetic segregation and by backcross is laborious and time consuming. We previously reported the development of the transgene killer CRISPR (TKC) technology that uses a pair of suicide genes to trigger self-elimination of the transgenes without compromising gene editing efficiency. The TKC technology enables isolation of transgene-free CRISPR-edited plants within a single generation, greatly accelerating crop improvements. Here, we presented two new TKC vectors that show great efficiency in both editing the target gene and in undergoing self-elimination of the transgenes. The new vectors replaced the CaMV35S promoter used in our previous TKC vector with two rice promoters to drive one of the suicide genes, providing advantages over our previous TKC vector under certain conditions. The vectors reported here offered more options and flexibility to conduct gene editing experiments in rice.     

Increasing selenium supply during the close-up dry period improves nutrient metabolism and attenuates inflammatory response after calving in dairy cows

The objective of this study was to investigate whether feeding selenium (Se)-replete cows a Se-yeast supplement in late pregnancy affects nutrient metabolism and inflammatory response during the periparturient period. Twenty cows were randomly assigned to two groups with 10 cows each. Cows in one group received Se-yeast at 0.3 mg Se/kg DM during the last 4 weeks before calving in addition to fed a TMR containing supplemented sodium selenite at 0.3 mg Se/kg DM (Se-yeast), while cows in another group were only fed a TMR containing supplemented sodium selenite at 0.3 mg Se/kg DM (Control). Blood samples were collected and analyzed for nonesterified fatty acids (NEFA), β-hydroxybutyrate (BHBA), glucose, insulin, tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, IL-6, serum amyloid A (SAA), haptoglobin (Hp), and albumin. In control cows, plasma NEFA, IL-1β, IL-6, SAA, and Hp levels increased after calving, but glucose, insulin, and albumin levels decreased after parturition. Se-yeast supplemental cows had lower postpartum concentrations of NEFA, TNF-α, IL-1β, IL-6, SAA, and Hp, and higher postpartum levels of glucose, insulin, and albumin compared with control cows. The results indicate that feeding Se-replete cows a Se-yeast supplement in late pregnancy improves nutrient metabolism and attenuates the inflammatory response after calving.     

Identification of Differentially Expressed Genes and Lipid Metabolism Signaling Pathways between Muscle and Fat Tissues in Broiler Chickens

In this study, signaling pathways and key differentially expressed genes (DEGs) involved in lipid metabolism in muscle and fat tissues were investigated. Muscle and abdominal fat tissues were obtained from 35-day-old female broilers for RNA sequencing. DEGs between muscle and fat tissues were identified. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses of DEGs were performed. A total of 6130 DEGs were identified to be significantly enriched in 365 GO terms, most of which were involved in biological processes, cellular components, and molecular functions in muscle and fat tissues. Three important lipid signaling pathways (pyruvate metabolism, the insulin signaling pathway, and the adipocytokine signaling pathway) were identified among the fat and muscle tissues of broilers. The key common DEGs in these pathways included phosphoenolpyruvate carboxykinase 2 (PCK2), acetyl-CoA carboxylase 1 alpha and beta (ACACA and ACACB), and the mitogen-activated protein kinase (AMPK) gene family. Hence, our findings revealed the pathways and key genes and gene families involved in the regulation of fat deposition in the muscle and fat tissues of broilers.     

丁酸钠对犊牛胃肠道发育的研究进展

丁酸钠是一种绿色新型饲料添加剂，目前在动物饲料中应用广泛，且效果良好。作者主要介绍了丁酸钠的生物学功能、对犊牛胃肠道发育的影响及其在犊牛日粮中的应用效果。重点总结了丁酸钠促进犊牛胃肠道发育的影响机制，包括作为胃肠道组织的能量来源调控激素和生长因子的分泌，提高消化和吸收相关酶的分泌及活性，提高肠道免疫细胞保护性蛋白的分泌等。现有研究表明，日粮干物质中添加0.3%丁酸钠就能达到促进犊牛生长和胃肠道发育的效果，但具体添加水平、添加阶段和方式等尚不明确。此外，日粮粗饲料水平、精料发酵所产生的内源丁酸对外源丁酸钠的作用效果是否有影响也鲜有报道，有待进一步开展研究。     

Effect of L-lysine Added in Diluent on the Fresh Semen Quality of Dorper Sheep

In order to explore the effect of different L-lysine levels added in semen dilution on the fresh Dorper sheep quality stored at liquid state,3 health Dorper ram were used to collect semen which were equal-packaged and added to solution containing different levels (0,0.1,0.2,0.3,0.4 and 0.5 g) L-lysine in 120 mL diluent and at 0,6,24,30,48,54,72 and 78 h,the sperm motility,membrane integrity and acrosome integrity were detected.The results showed that the sperm motility in groups added 0,0.1 and 0.2 g L-lysine were higher than other groups,and that in 0.4 and 0.5 g L-lysine groups decreased faster than other groups;The sperm motility in 0.5 g L-lysine group reduced to 0 at 30 h,and it reduced to 0 at 48 h in the group added 0.4 g L-lysine;The effective survival time and survival index in the group added 0.1 g L-lysine was higher than other groups (P< 0.05);The membrane integrity of 0.1 g group was the highest,that had no significant difference with control group from 0 to 54 h (P >0.05),while after 72 h the two groups had significant differences (P< 0.05).The membrane integrity in 0.4 and 0.5 g groups were the worst,and that was significant different with other groups (P< 0.05);The acrosome integrity of 0.1 g group was the highest which had no significant difference with control group (P >0.05),0.4 and 0.5 g groups were the worst,the difference between them was not significant (P >0.05),while was extremely significant difference with other groups except for 0 and 6 h (P< 0.01).The results suggested that dilution added a high concentration of L-lysine could inhibit sperm quality,when the count of L-lysine was more than 0.2 g,sperm quality was significantly decreased,adding 0.1 g L-lysine could improve Dorper sheep fresh sperm quality stored at liquid state.     

Symbiotic N<Subscript>2</Subscript> fixation and nitrate utilisation in irrigated lucerne (<Emphasis Type="Italic">Medicago sativa</Emphasis>) systems

Symbiotic N₂ fixation by lucerne (Medicago sativa) has capacity to provide significant inputs of N to agro-ecosystems, and the species has also been shown to scavenge soil mineral N and thus act as a sink for excess reactive N. The balance between these two N cycle processes was investigated in an extensive irrigated lucerne growing region where nitrate contamination of groundwater has been reported. We sampled 18 permanent pure lucerne stands under irrigation for standing dry matter, total shoot N, and N₂ fixation using ¹⁵N natural abundance along with activity of the inducible enzyme nitrate reductase as indicators of use of soil NO₃⁻ by lucerne. On average 65% of lucerne N was obtained from symbiotic N₂ fixation. Converting standing dry matter estimates to annual N₂ fixation amounts we calculated average N₂ fixation of 311 kg N/ha, including N in roots and nodules. Uptake of N from soil by lucerne was calculated to be 181 kg N/ha/year. We were not able to identify the source of this soil mineral N, although nitrate reductase activity of lucerne was higher than that of non-N₂ fixing species examined.     

棉铃发育过程中不同组织IAA含量变化特点

为了解IAA与棉纤维品质形成间的关系及棉铃各部位IAA积累转运特点,本研究分析了棉铃各部分在发育过程中IAA含量的变化特点。通过选择陆地棉标准系TM-1、长纤维种质‘母35’和短纤维种质‘208’3个纤维长度差异较大的陆地棉材料,利用酶联免疫法对浙江杭州、海南三亚两地田间试验的各材料不同发育阶段的棉仁、棉籽壳、棉纤维、棉铃壳进行了IAA含量的测定。结果表明,不同参试材料的棉仁IAA含量存在明显差异,棉籽壳、棉纤维与棉铃壳的IAA含量峰值出现时间晚于棉仁,棉铃各个组织IAA含量具有随棉铃组织从内向外逐渐降低的趋势。研究表明,IAA可能是棉纤维发育的正效应因子,且棉铃发育过程中IAA可能呈初期在棉仁富集、随发育进程沿棉铃组织由内向外运输的规律。     

外源没食子酸丙酯对番茄幼苗响应低温胁迫的生理特性影响

以‘金鹏10号’番茄幼苗为试材,研究了外源没食子酸丙酯(PG)对低温逆境下番茄幼苗生理特性的影响。结果表明:外源PG诱导能明显提高番茄幼苗的抗冷性;与低温对照相比,经PG诱导的番茄幼苗叶片的相对电导率和丙二醛(MDA)含量显著降低,过氧化物酶(POD)和超氧化物歧化酶(SOD)活性、叶绿素、脯氨酸和可溶性糖含量显著提高。其中,PG浓度为2.0mmol/L的处理表现最佳。