Embryo-mediated genome editing for accelerated genetic improvement of livestock

Selecting beneficial DNA variants is the main goal of animal breeding. However, this process is inherently inefficient because each animal only carries a fraction of all desirable variants. Genome editing technology with its ability to directly introduce beneficial sequence variants offers new opportunities to modernize animal breeding by overcoming this biological limitation and accelerating genetic gains. To realize rapid genetic gain, precise edits need to be introduced into genomically-selected embryos, which minimizes the genetic lag. However, embryo-mediated precision editing by homology-directed repair (HDR) mechanisms is currently an inefficient process that often produces mosaic embryos and greatly limits the numbers of available edited embryos. This review provides a summary of genome editing in bovine embryos and proposes an embryo-mediated accelerated breeding scheme that overcomes the present efficiency limitations of HDR editing in bovine embryos. It integrates embryo-based genomic selection with precise multi-editing and uses embryonic cloning with elite edited blastomeres or embryonic pluripotent stem cells to resolve mosaicism, enable multiplex editing and multiply rare elite genotypes. Such a breeding strategy would enable a more targeted, accelerated approach for livestock improvement that allows stacking of beneficial variants, even including novel traits from outside the breeding population, in the most recent elite genetic background, essentially within a single generation.     

Phenolic extract of Morchella angusticeps peck inhibited the proliferation of HepG2 cells in vitro by inducing the signal transduction pathway of p38/MAPK

Morchella angusticeps Peck, one of the most popular edible mushrooms, has attracted great attention due to its delicious taste and healthy properties. However, both its biological effects and the possible mechanism of action have not yet been known. We investigated the anti-proliferative activity of the phenolic extract derived from Morchella angusticeps Peck (MPE) against HepG2 human hepatocellular carcinoma cells. Results showed that MPE at non-cytotoxicity doses significantly inhibited the proliferation of HepG2 cells in a dose-dependent manner with inhibitory rates ranging from 18 to 90% (P<0.01). The possible mechanism might be that MPE induced apoptosis through initiating the mitochondrial death pathway by regulating Bax, Bcl-2 and cleaved caspase-3. On the other hand, MPE might trigger cell cycle arrest at G₀/G₁/S phases by managing p21, Cyclin D1, cyclin-dependent kinases-4 (CDK4) and proliferating cell nuclear antigen (PCNA). Additionally, MPE downregulated TRAF-2 and p-p53, while upregulated p-ASK1 and p-p38. Therefore, it could be inferred that MPE might induce the anti-proliferative function to HepG2 cells through the p38/MAPK signal transduction pathway.     

槲树叶提取液对冷藏期间桃品质及抗氧化系统的影响

以生理成熟期‘新泰红’桃果实为试材,分别用0、2.5、5.0、7.5 g/L槲树叶提取液浸泡果实,以蒸馏水为对照,并于0℃冷藏,测定冷藏期间果实可溶性蛋白等品质指标以及酶活力与活性氧等物质,探讨槲树叶提取液对采后桃果实贮藏品质的影响。结果表明:槲树叶提取液处理能够有效延缓冷藏期间桃果实冷害指数升高以及硬度下降,使整个冷藏期间果实可溶性蛋白、可溶性糖、维生素C含量维持相对较高水平,并且能够有效延缓膜脂过氧化产物(丙二醛)以及活性氧物质(超氧阴离子自由基、过氧化氢和羟基自由基)积累;同时,槲树叶提取液处理还能够显著抑制果实多酚氧化酶(PPO)、苯丙氨酸解氨酶(PAL)活性,从而延缓冷藏期间桃果实褐变程度。综上所述,槲树叶提取液处理能够缓解冷藏期间桃果实冷害症状,以5.0 g/L槲树叶提取液处理效果最为明显。     

土霉素对柑橘黄龙病的防治效果及对柑橘产量和品质的影响

为有效防控柑橘黄龙病,于2017年在佛罗里达大学柑橘研究与教育中心的奥本代尔市柑橘试验场进行田间试验筛选黄龙病的防治药剂及其浓度,测定注射土霉素后叶片中柑橘黄龙病菌亚洲种Cadidatus Liberibacter asiaticus、土霉素、淀粉含量、柑橘产量、出汁率、可溶性固形物含量和酸度,显微镜下观察注射土霉素后淀粉粒的分布。结果表明,浓度为1.6g/株土霉素处理180 d后柑橘叶片中黄龙病菌亚洲种含量减少幅度最大,为91.78%;注射浓度为1.6g/株土霉素后7d,叶片中土霉素含量达142.2 μg/kg,注射后11 d叶片中土霉素含量达到最大,为239.8 μg/kg,注射后45 d叶片中土霉素含量低至99.6 μg/kg以下;注射后7~90d,叶片中黄龙病菌亚洲种含量呈波浪形变化,注射90 d后叶片中黄龙病菌亚洲种含量一直处于增长趋势,叶片中黄龙病菌亚洲种含量总体随着土霉素含量的升高而降低;注射浓度为1.6g/株土霉素后,叶片中淀粉含量大幅度下降,60d时达到最低值,为3.3 μg/mm²,90d时达到峰值,为14.5 μg/mm²;单株产量为16.7 kg,与注射浓度为0.8 g/株土霉素处理差异不显著,但均显著高于其它2个处理;柑橘出汁率、可溶性固形物和酸度均与清水对照差异不显著。表明土霉素可有效抑制黄龙病菌亚洲种,减少柑橘叶片内淀粉含量,增加柑橘产量,但对果实品质未产生显著影响。     

Crude garlic extract significantly inhibits replication of grapevine viruses

Efforts to control viral diseases of grapevine include the production of certified material and development of virus-resistant transgenic grapevines. However, effective antiviral agents, once the viruses have infected the plants, are still lacking. This study shows that a crude garlic extract has significant antiviral activity against grapevine viruses. Replication of grapevine leafroll-associated virus 2 (GLRaV-2) was obviously inhibited in grapevine cv. Cabernet Sauvignon calli treated with diluted (1:100) garlic extract. The relative RNA levels of GLRaV-2 and grapevine fleck virus (GFkV) in cv. Summer Black grapevine in in vitro-grown plantlets 10 days after treatment with diluted (1:100) garlic extract were about 22% and 20%, respectively, of that in controls. The viral RNA accumulation of GLRaV-2, GFkV, grapevine virus A (GVA), grapevine fanleaf virus (GFLV) and grapevine rupestris stem pitting-associated virus (GRSPaV) in field-grown grapevine cv. Centennial Seedless plants sprayed with diluted (1:100) garlic extract were about 31–40%, 26–38%, 18–31%, 17–42% and 15–18%, respectively, of that in controls. Moreover, the garlic extract treatment led to a significant decrease in viral RNA accumulation of GLRaV-3, GLRaV-2, GVA, GFkV, GFLV, GRSPaV and grapevine Pinot Gris virus in pot-grown grapevine cv. Shine Muscat plants, and viral disease symptoms in these plants were obviously attenuated. In addition, this extract significantly induced expression of pathogenesis-related protein genes and stimulated activity of antioxidant enzymes in grapevines. Taken together, these results indicate that the crude garlic extract acts as a significant inhibitor against a broad range of grapevine viruses.     

探究姜和大蒜粗提取液对甘蓝枯萎病菌的抑制作用

研究旨在了解不同浓度姜和大蒜粗提取液对甘蓝枯萎病菌的抑制效果及对菌丝形态结构的影响,以此初步获得最适合的抑制病原菌的植物粗提取液浓度。以姜和大蒜粗提取液为供试材料,测定不同浓度2种粗提取液对甘蓝枯萎病菌尖孢镰刀菌的生长情况、孢子浓度、孢子萌发率及菌丝结构的影响。结果表明,2种粗提取液均能抑制并降低尖孢镰刀菌的生长速率、孢子浓度和孢子萌发率,且大蒜粗提取液比姜粗提取液抑制效果更显著,在低浓度下对菌丝形态结构的影响也比姜粗提取液明显,大蒜粗提取液在各浓度下与对照相比均达极显著差异,且大蒜浓度为0.4 g/mL时对尖孢镰刀菌孢子萌发率抑制达100%。该试验证实大蒜对甘蓝枯萎病菌有极显著抑制作用,为使用间种或轮作大蒜进行田间防治甘蓝枯萎病菌提供了参考。     

生物炭浸提液对小白菜种子萌发及幼苗荧光作用的影响

生物炭被广泛应用于低产地改良以及重金属污染土壤修复,但是其应用可能对植物存在潜在毒性。为了揭示生物炭的潜在植物毒性,用不同来源、不同浓度生物炭浸提液对小白菜种子进行处理,检测种子萌发以及幼苗生长相关指标。结果显示,酒糟生物炭和梨木生物炭浸提液对小白菜种子萌发以及幼苗生长具有明显作用,但不同生物炭浸提液以及不同浓度浸提液影响差异较大。①酒糟生物炭和梨木生物炭浸提液原液均显著降低小白菜种子萌发(P<0.05),但稀释后的浸提液却促进小白菜种子萌发,尤其是稀释50倍后梨木炭浸提液和酒糟炭浸提液处理的小白菜种子萌发率分别比对照高16.67%和8.33%;②生物炭浸提液可提高小白菜幼苗株高和根系活力,尤其浸提液原液处理的植株株高、根系活力显著高于对照(P<0.05);酒糟生物炭浸提液的促进作用明显低于梨木生物炭;③生物炭浸提液均可提高小白菜幼苗叶片叶绿素含量、光化学效率（Fv/Fm）、实际量子产量（ΦPSⅡ）和表观光电子传递速率（ETR）等,且供试植株幼苗叶片叶绿素含量、Fv/Fm、ΦPSⅡ、ETR等随生物炭浸提液浓度增加而增加;但酒糟浸提液对小白菜幼苗叶片荧光作用的影响低于梨木生物炭浸提液;④生物炭浸提液能显著降低幼苗丙二醛含量 (P<0.05),且其随生物炭浸提液浓度增加而降低。由此表明,酒糟生物炭和梨木生物炭浸提稀释液均可提高小白菜种子萌发率及幼苗根系活力,增强幼苗叶片的荧光作用,但酒糟生物炭浸提液的影响作用低于梨木生物炭。     

天麻茎秆水提物的抗惊厥作用

【目的】研究天麻茎秆(Gastrodia elata stalk)水提物的抗惊厥作用及其可能的作用机制。【方法】以SPF级ICR小鼠为研究对象,采用戊四氮(PTZ)和N-甲基-D -冬氨酸(NMDA)构建2种小鼠癫痫模型,以卡马西平(80 mg/kg)为阳性对照,以相同体积的蒸馏水为空白对照,通过检测小鼠脑中γ-氨基丁酸(GABA)、谷氨酸(Glu)、一氧化氮(NO)、五羟色胺(5-HT)、Na⁺-K⁺-ATP酶(Na⁺-K⁺-ATP)、天冬氨酸(Asp)、总抗氧化能力(AOC)、过氧化氢酶(CAT)、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)、谷胱甘肽还原酶(GSR)、丙二醛 (MDA)、白细胞介素1β(IL-1β)、白细胞介素10(IL-10)、肿瘤坏死因子α(TNF-α)水平的变化,分析天麻茎秆水提物高、中、低(2 000,1 000,500 mg/kg) 3个剂量对模型小鼠惊厥发作的行为学影响,观察各组小鼠脑皮层及海马组织的病理学变化,免疫组织化学方法测定脑皮层、海马组织中c-Fos、TLR4蛋白的表达水平。【结果】与空白对照组相比,2种模型组小鼠发生惊厥数及死亡率极高,其脑组织中GABA、5-HT、Na⁺-K⁺-ATP、AOC、CAT、GSH、SOD、GSR、IL-10的含量降低,Glu、Asp、NO、MDA、IL-1β、TNF-α的含量升高,脑皮层和海马组织中c-Fos、TLR4蛋白表达水平升高,海马神经元受损。天麻茎秆高、中、低剂量组和卡马西平组均可改善由PTZ、NMDA所致上述指标的变化,延长小鼠的惊厥潜伏期,缩短惊厥持续时间,减少惊厥次数,降低惊厥强度及死亡率。【结论】天麻茎秆具有良好的抗惊厥作用,能缓解由PTZ和NMDA诱发的小鼠惊厥。     

膜荚黄芪根水浸液对4种作物种子萌发及幼苗生长的影响

通过室内生物测定法,研究了黄芪根水浸液对4种作物种子萌发及幼苗生长的化感影响。结果表明,黄芪根水浸液对4种作物种子萌发及幼苗生长均有明显的化感作用,其中根受到的化感抑制大于地上部分,种子萌发阶段受到的化感抑制大于幼苗生长阶段。不同受体的发芽势、最长根长和根冠比受到的化感抑制最强。4种作物的化感敏感性为小麦>板蓝根>大麦>大豆。因此,选择与黄芪轮作、套作的作物时,优先选择大豆和大麦,其次为板蓝根。     

中草药复配液在脐橙保鲜中的应用研究

以赣南脐橙为试材,研究了肉桂、丁香、丹皮、细辛、五倍子五种中草药提取液(浓度为1 g/mL)中的任意三种以1∶1∶1比例形成的10种复配液对意大利青霉和指状青霉的抑制效果以及效果最佳的复配液对脐橙的保鲜效果。结果表明,复配液CM-7(丁香、细辛、丹皮)对指状青霉和意大利青霉的抑菌圈直径分别为40.97 mm和39.03 mm,抑菌效果最好。对脐橙保鲜效果表明,与对照组相比,在贮藏中后期(40～90 d),CM-7处理降低了果实的腐烂率和失重率,延缓了VC和总糖含量的降解,保持了果实良好的营养品质。