Development of a kompetitive allele‐specific PCR marker for selection of the mutated Wx‐D1d allele in wheat breeding

Waxy (Wx) protein is a key enzyme for synthesis of amylose in endosperm. Amylose content in wheat grain influences the quality of end‐use products. Seven alleles have been described at the Wx‐D1 locus, but only two of them (Wx‐D1b, Wx‐D1e) were genotyped with codominant markers. The waxy wheat line K107Wx1 developed by treating ‘Kanto 107’ seeds with ethyl methanesulphonate carries the Wx‐D1d allele. However, no molecular basis supports this nomenclature. In the present study, DNA sequence analysis confirmed that a single nucleotide polymorphism in the sixth exon of Wx‐D1 changed tryptophan at position 301 into a termination codon. Based on this sequence variation, a PCR‐based KASP marker was developed to detect this point mutation using 68 BC₈F₁ plants and 297 BC₈F₂ lines derived from the cross ‘Ningmai 14’*9/K107Wx1. Combined with codominant markers for the Wx‐A1 and Wx‐B1 alleles, waxy and non‐waxy near‐isogenic lines were distinguished. The KASP marker was efficient in identifying the mutant allele and can be used to transfer waxiness to elite lines.     

利用钾吸收基因表达评价葡萄叶面喷施钾肥效果和喷施浓度

【目的】叶面喷施钾肥可以快速、 高效地为葡萄补充钾营养,促进葡萄的高产和优质,已经被广泛应用于葡萄生产中。本实验通过对葡萄叶面喷施不同种类及浓度钾肥,测定葡萄叶片和果实等生理指标变化,以及钾吸收相关基因的表达变化,从生理和基因水平上评价这些钾肥的喷施效果,为葡萄生产中钾肥的施用提供一定指导。【方法】本实验以‘夏黑’葡萄为试材,选择两个葡萄生长关键时期盛花期和果实膨大期分别对葡萄叶片喷施0.2%、 0.5%和0.8%三种浓度的K2SO4、 K2CO3、 K2SO4·2MgSO4和KCl。然后对葡萄叶片和新梢的生长率,坐果率,叶绿素含量,单粒重,可溶性固形物等生理指标进行统计分析,并利用荧光定量PCR技术分析4个钾吸收相关基因VvHAK13、 VvKEA2、 VvSIRK和VvSORK的表达情况。【结果】叶面喷施4种钾肥后,葡萄叶片和新梢的生长率,坐果率,叶绿素含量,单粒重,可溶性固形物等各项生理指标均有不同程度的提升。钾肥种类不同,最适喷施浓度不同,同种钾肥在葡萄盛花期和果实膨大期的最适喷施浓度也有所不同,四种钾肥在果实膨大期的最适喷施浓度普遍高于盛花期。四个钾吸收相关基因在喷施不同种类及浓度钾肥后也表现出不同的表达模式,总体来讲VvKEA2、 VvSIRK 、 VvSORK的表达上调,而VvHAK13的表达下调。果实膨大期,需喷施较高浓度的钾肥,钾吸收相关基因才表现出较强烈的响应,而在盛花期则只需喷施较低浓度的钾肥。综合生理指标和基因表达两方面结果,得出4种钾肥效果依次为K2SO4·2MgSO4＞K2SO4＞K2CO3＞KCl。盛花期K2SO4和K2CO3的最适喷施浓度为0.5%,K2SO4·2MgSO4和KCl的最适喷施浓度为0.2%; 果实膨大期K2SO4、 K2SO4·2MgSO4和KCl的最适喷施浓度为0.5%,K2CO3的最适喷施浓度为0.8%。【结论】葡萄叶面喷施钾肥可以有效促进葡萄叶片和果实的生长发育,四种钾肥的效果依此为: K2SO4·2MgSO4＞K2SO4＞K2CO3＞KCl。盛花期K2SO4和K2CO3的最适喷施浓度为0.5%,K2SO4·2MgSO4和KCl的最适喷施浓度为0.2%; 果实膨大期K2SO4、 K2SO4·2MgSO4和KCl的最适喷施浓度为0.5%,K2CO3的最适喷施浓度为0.8%。适宜的喷施浓度可以有效提高钾吸收相关基因的表达,是其提高钾吸收利用的机理之一。     

激素配比和pH值对蓝莓试管苗增殖生长的影响

该文以对土壤p H值适应性截然不同的蓝莓品种‘斯巴坦’和‘北陆’试管苗为试材,研究二者组织培养条件下对激素配比和p H值的要求。结果表明:‘斯巴坦’试管苗增殖培养基最佳激素配比为ZT0.3 mg/L+IBA 0.2 mg/L,p H值4.5;‘北陆’试管苗增殖培养基最佳激素配比ZT 0.3 mg/L+IBA 0.1 mg/L,p H值5.0。     

直剪法用于糙米内摩擦角的测定与研究

利用土工合成材料综合测定仪（直剪仪）,以水分含量14.57%的糙米为样品,测定了在不同的法向压应力、不同的剪切速率下糙米的剪切应力并计算出内摩擦角。实验结果表明：糙米的剪切应力随着法向压应力（25kPa～100kPa）的增大而增大;剪切速率（1.33mm/min～2.67mm/min）较低时糙米的内摩擦角随着法向压应力的增大而减小;随着剪切速率（1.33mm/min～5.20mm/min）的增大,糙米的内摩擦角减小。     

基于纳米碳添加的减量施肥模式对寒地水稻产量及氮肥效率的影响

为解决减量施肥造成北方寒地水稻肥料供给不足和产量低的问题,以纳米碳增效剂为材料,采用仪器快速检测和植株常规测定分析方法,研究纳米碳添加的常规施肥和减量施肥模式对水稻产量性状和肥料利用效率的影响。结果表明：与常规施肥(CK)相比,纳米碳添加的常规施肥模式(SM-A)、减施氮肥10%(SM-B)和减施氮肥20%(SM-C)对水稻植株形态特征、光合生理特性、产量性状和肥料利用效率产生显著的影响,其中以SM-B增幅最大,SM-A次之,SM-C最小;SM-A、SM-B和SM-C增加经济效益分别为563.56、1374.20和-1391.08元/hm ²,以SM-B增效值最高。本研究表明,SM-B能改善生育期光合生理特性,提高植株产量和肥料利用效率,是寒地水稻较为理想的减量施肥安全利用模式。     

硫酸蒽酮法测定柑橘属植物多糖的研究

采取浸提法从丑橘、甜橙、甜心柚、血橙、芦柑5种柑橘属果皮中提取多糖并测定其含量。结果表明,血橙果皮中含糖量在5种果皮中最低,其次为甜橙果皮、甜心柚果皮、芦柑果皮,丑橘果皮中含糖量在5种果皮中最高。甜橙果皮和甜心柚果皮含糖量差距不大。     

Development and application of a molecular marker for TMV resistance based on <Emphasis Type="Italic">N</Emphasis> gene in tobacco (<Emphasis Type="Italic">Nicotiana tabacum</Emphasis>)

Tobacco mosaic virus (TMV) caused serious loss in yield and quality of tobacco every year. It is a long-term goal to improve the tobacco resistance against TMV by tobacco breeding. N gene was the firstly reported TMV-resistant gene, which showed resistance against all Tobamoviruses except the Ob stain and belonged to the toll-interleukin-1 receptor/nucleotide-binding site/leucine-rich repeat class of plant resistance (R) genes. At present, N gene had already been widely used in tobacco conventional breeding, but there is rare available molecular maker used in marker-assisted selection of TMV resistance. In this study, we designed a pair of primers that specific amplify N gene fragment based on the sequence of N gene intron III, named N-marker. Then, we identified TMV resistance by two selecting methods, PCR with N-marker and inoculated with the TMV-C strain. Results from the two method showed that (1) 13 varieties among 67 tobacco varieties displayed hypersensitive reaction when inoculated with the TMV-C strain, also contained N gene fragments screened by PCR with N-marker; (2) 105 strains of 200 BC1 strains showed resistance against TMV when inoculated with TMV-C strain, meanwhile, 103 of the 105 strains contained N gene fragment verified by PCR with N-marker. Therefore, the N-marker is reliable for high throughput screening of germplasm resources and tobacco breeding materials in selection of N-mediated TMV resistance. Our study not only developed a molecular marker for tobacco breeding, but also identified new germplasm resources that are resistant to TMV.     

基于EDEM的猪粪接触参数标定

为准确快速获得畜禽粪便的接触参数,该研究通过物理堆积试验与仿真方法对猪粪接触参数进行了标定。测定了不同含水率下猪粪的堆积角,建立了含水率与堆积角的回归方程;基于Hertz-Mindlin with JKR球体粘结模型,进行了离散元仿真模拟;采用筛选试验设计(Plackett-BurmanDesign,P-BD)对10个初始参数进行了筛选,发现JKR(Johnso-Kendall-Roberts)表面能、颗粒间滚动摩擦系数、颗粒间碰撞恢复系数对猪粪堆积角影响显著;并根据响应曲面试验设计(Box-BehnkenDesign,B-BD)建立了堆积角与显著性参数的二阶回归模型,得到了3个显著性参数值分别为JKR表面能0.03J/m~2、颗粒间滚动摩擦系数0.27、颗粒间碰撞恢复系数0.54;将仿真所得堆积角与物理试验值进行对比验证,相对误差为4.27%。结果表明,该研究提出的标定方法能准确模拟物理堆积试验,可为畜禽粪便接触参数的标定提供参考。     

浅谈猪精液保存在人工授精中的发展与展望

在世界上主要的猪肉生产国中,猪人工授精(AI)技术的应用率在90%以上。尽管猪冷冻精液技术在不断进步,但由于其效率不高,在猪场实际生产中并不多见使用。在猪人工授精技术中,液态精液是目前应用最广泛的精液,与传统的牛冷冻精液不同,其独特的精子微胶囊保护膜技术,可以对猪精子进行更加完善的液态保存,因此液态精液是目前猪人工授精使用的主要方式。文章综述了猪精液保存技术在人工授精中的应用,并展望了今后的发展方向,为人工授精技术助力恢复生猪生产提供参考。