河南省花生地方资源蛋白质和脂肪含量分析及其育种利用策略

对233份河南省地方花生资源进行了蛋白质含量、含油量、油酸和亚油酸含量的全面测定,并与省外和国外资源的相关性状进行了比较分析。在河南地方品种资源中,蛋白质含量中等,平均含油量和油酸含量相对较高,但缺乏蛋白质含量超过30%或含油量超过56%、油酸含量超过70%的突出材料。河南省目前高油品种选育有明显进展,育成了一批高油花生品种,但育成品种蛋白质含量普遍偏低。提出了充分利用现有地方品种资源,积极采用远缘杂交、诱变、分子标记辅助选择技术及现代基因工程技术创制优良种质,选育优质专用品种的育种策略。     

花生籽仁外观和营养品质特征及食用型花生育种利用分析

花生籽仁的外观品质和营养品质是评价食用花生品种的重要指标。本研究对285个不同类型或来源的花生种质资源进行了籽仁外观和营养品质性状的检测和分析,旨在为食用型花生品种选育提供依据。按照植物学类型分析了各类型的8个外观性状和5个营养品质性状的分布。结果表明,我国花生种质资源包含丰富的外观性状变异类型,能够满足不同加工用途对原料的需求,但总体上高油酸、高蛋白和低脂肪资源类型偏少。借鉴美国等烤制花生仁和糖果花生的外观和营养品质特征及我国煮食和烤制花生的生产消费市场,提出了食用型品种选育亲本的策略和备选品种类型,烤制和糖果用途花生育种亲本选配应以普通型、中间型或珍珠豆型中粒农家品种或育成品种为骨干亲本,进一步改良高油酸、高蛋白和低脂肪等品质性状。     

辽宁花生品种系谱分析及农艺性状的演变

分析了辽宁省1949-2012年育成的95个花生品种系谱及农艺性状的演变。结果表明:辽宁花生育成品种共涉及100个亲本,其中,来自辽宁的有45个,49个是育种单位的中间材料,鲁花12号、白沙1016、伏花生、豫花11号等是辽宁花生育成品种的骨干亲本。进入21世纪以来,辽宁省育成的花生品种株高、侧枝长逐渐增加,从变异区间来看,百仁重、出米率呈增加趋势,而粗蛋白与粗脂肪含量变化较小。在分析辽宁省花生育种背景的基础上,提出辽宁省花生育种上宜重视回交手段的利用,发展食用型品种,把抗旱、抗寒、抗病、抗虫、耐连作作为重要的育种目标,利用生物技术手段和野生资源加速育种进程,进一步拓宽辽宁花生品种的遗传基础。     

高油酸油菜遗传育种研究进展

油酸（C18∶1）是双低菜籽油的主要脂肪酸之一,合理的高油酸菜籽油脂肪酸组分更有利于人体健康,因此提高油酸含量是双低油菜品质育种的一个重要方向。当前我国高油酸油菜品种选育相关研究进展缓慢,高油酸菜籽油产业化进程急需提速。围绕高油酸油菜主要从四个方面开展论述：国内外利用理化方法成功创建的油菜高油酸种质资源及其高油酸性状遗传模式;油菜高油酸性状的控制基因及其突变位点;世界上高油酸品种的培育以及我国高油酸品种的发展趋势;当前高油酸品种的不足及今后高油酸油菜品种的改良途径。为油菜育种工作者较全面地展示了国际上高油酸油菜在遗传育种方面的研究成果,也为今后我国高油酸油菜的进一步发展提供了参考。     

高油酸花生新品种宇花91的选育

宇花91是青岛农业大学选育的高油酸花生新品种。以普通油酸含量品种鲁花11号为母本,F435型高油酸花生品种开农1715为父本配置杂交组合。利用PCR产物测序法筛选获得F_1代真杂种,对F_2代单株提取叶片基因组DNA,利用PCR产物测序法筛选基因型纯合的单株个体。对当代收获的单株籽粒利用近红外法多粒模型测定油酸、亚油酸含量,筛选油酸含量在80%以上且油酸亚油酸比值在10.0以上的单株种植成株行,随后利用系谱法进行选择育种。宇花91荚果为普通型小果,网纹较细、较明显,百果重148.06 g,百仁重63.31 g,果皮薄,出米率75.15%。籽仁长椭圆形,种皮粉红色、无裂纹,内种皮白色。籽仁蛋白质含量26.57%,脂肪含量52.72%,油酸含量80.40%,亚油酸含量2.50%,棕榈酸含量5.57%,油酸亚油酸比值32.16。苗期生长旺盛,封垄早,结果集中,中抗叶斑病和青枯病。2017年参加山东省夏播多点试验,平均荚果产量215.79 kg/667 m~2,比对照花育20号增产15.27%;平均籽仁产量157.33kg/667m~2,比对照花育20号增产21.64%。2018年通过国家花生品种登记,登记号:GPD花生(2018) 370210,适于在山东花生产区种植。     

不同温度型花生品种的抗衰老特性

以我国北方4个大花生品种为研究材料,于花生结荚期到收获,用红外测温仪对各品种的冠层温度进行连续观测,并测定了花生主茎功能叶片中叶绿素、可溶性糖、可溶性蛋白、丙二醛（MDA）含量及硝酸还原酶（NR）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性。结果表明,不同品种花生冠层温度存在明显差异。冠层温度低的品种比冠层温度高的品种叶绿素、可溶性糖、可溶性蛋白含量及NR、SOD和CAT活性明显偏高,而MDA含量明显偏低,且越往生育后期越明显,说明冠层温度低的品种比冠层温度高的品种生活力更旺盛,抵抗衰老能力更强。因此,冠层温度可作为花生抗衰老能力的一个重要指标,用于指导花生育种和栽培等生产实践。     

中国主要花生品种品质性状关联分析

以来自全国的136个花生主要育成品种及育种骨干亲本作为供试群体,测定其蛋白质含量、脂肪及油酸含量等性状。选用64个均匀分布在复合遗传图谱不同遗传连锁群上多态性较好的SSR标记进行多位点扫描。通过标记的基因型值,利用Structure软件对群体进行结构划分并得到结构划分的矫正Q值,采用Tassel 软件中GLM（Q）方法将供试花生品种连续3年的品质性状与SSR标记进行关联分析。结果表明：①依据基因型数据对材料群的结构划分,供试群体最终可被划分为5个亚群,依据群体特点和结构分析可以表明供试的花生材料是适合关联分析的;②通过关联分析,共发掘与2010年、2011年、2012年品质性状显著关联的SSR位点分别有18个、31个、26个;③通过综合分析,能够连续3年重复检测出与品质性状关联的SSR位点4个,共计等位变异位点40个。     

花生抗青枯病种质脂肪酸组成的遗传多样性

通过对123份不同类型抗青枯病花生种质种子脂肪酸的鉴定测试,分析了抗青枯病花生种质在这些性状方面的遗传分化,并与6006份资源组成的花生基础收集品进行了比较。研究结果表明,我国抗青枯病花生资源的油酸含量平均为51.78%,显著高于基础收集品的对应值(45.64%);亚油酸含量平均为28.88%,显著低于基础收集品的对应值(34.36%);高油酸种质较多,油酸含量达61%以上的资源23份,所占比重为18.7%,显著高于基础品中的相应比重(2.65%)。标准差、变异系数以及遗传多样性指数的分析结果表明,抗青枯病资源在油酸和亚油酸含量方面的遗传分化程度高。     

河南省花生农家品种资源农艺和品质性状分析

综合分析河南省花生农家品种资源的农艺和品质性状,为花生的遗传育种提供理论依据。以128个不同地域来源的河南省农家品种为材料,田间调查株型、分枝型和开花习性等植物学性状,收获考种测定主茎高、侧枝长、总分枝数、结果枝数、单株产量、百果重和百仁重农艺性状,并测定蛋白质、脂肪、油酸和亚油酸含量。结果表明,农家品种以密枝型为主,农艺性状中百仁重的变异系数最大,为31.1%,其次为单株产量和总分枝数,分别为27.5%和22.2%,变异系数最小的为侧枝长,为12.5%;品质性状方面,河南农家品种资源的脂肪含量较高,脂肪含量55%以上的有10个花生品种。蛋白质含量偏低,最高仅25.3%,油酸含量中等,平均45.4%,最高的52.4%。本研究表明河南省农家品种的农艺性状表现丰富的遗传多样性,品质方面脂肪含量相对较高,合理利用河南省农家品种资源,可为花生品质改良提供优质性状的亲本。     

中国花生核心种质中高油酸材料的分布和遗传多样性

利用代表花生基础资源的核心种质分析花生高油酸资源的分布和遗传多样性,结果表明：在花生核心种质中油酸含量高于57%的种质40份,主要分布在密枝亚种（普通型25份和龙生型8份）,少数分布在疏枝亚种（珍珠豆型6份和中间型1份）; 除了10份资源来源于国外（ICRISAT 7 份,美国1份,日本1份和韩国1份,其他种质资源来源于中国12个省市。 同时发现高油酸种质中3份资源的含油量在55%左右,分别是Zh.h4094（油酸66.70%,含油量54.99%）, Zh.h4029（油酸63.50%,含油量55.58%）和Zh.h4319（油酸59.70%,含油量56.04%; 小区产量超过3000 kg/ha 有10 份种质,前三位分别是Zh.h0883 （4086.06kg/ha）, Zh.h1182（3955.00kg/ha）和 Zh.h2910 （3741.00kg/ha）。基于植物学和产量性状分析,前5个主成份（PC）可以解释81.17% 的变异。聚类分析,在域值为0.1942时,可分为6个组。 因此中国花生核心种质中高油酸种质存在丰富的遗传多样性,而且分布较广,高油酸种质的获得对花生高油酸育种提供基础材料。     

Hemodynamic effects of oleic acid in newborn lambs: role of arachidonic acid metabolites.

Oleic acid injection produces acute lung injury and pulmonary hypertension in adult animals. In other types of acute lung injury, such as that caused by E. coli endotoxin, metabolites of arachidonic acid are important mediators of pulmonary hypertension. In order to understand the hemodynamic response of newborn animals to oleic acid injection and the contribution of arachidonic acid metabolites to that response, we injected oleic acid into awake, chronically instrumented newborn lambs. The hemodynamic response of lambs to injections of oleic acid alone was compared to their response after pretreatment with either FPL57231, a putative leukotriene receptor antagonist, or indomethacin, a cyclooxygenase synthesis inhibitor. Oleic acid caused acute pulmonary hypertension associated with an increase in protein-rich lung lymph fluid. Systemic hemodynamic effects were variable. FPL57231 completely blocked the oleic acid-induced pulmonary hypertension while indomethacin significantly attenuated the response. Therefore, metabolites of arachidonic acid metabolism appear to be important mediators of oleic acid-induced pulmonary hypertension in newborn lambs.     

国际半干旱热带地区作物研究所花生微核心种质含油量及脂肪酸分析与鉴定

以国际半干旱热带地区作物研究所(ICRISAT)花生微核心种质为材料,系统分析测试含油量和脂肪酸组成。分析结果表明,ICRISAT花生微核心种质的含油量平均为51.67%,变异范围49.16%~55.44%,珍珠豆型资源的含油量高于其他类型,发掘出高油种质1份。在主要脂肪酸中,棕榈酸平均含量10.74%,变异范围7.9%~13.5%;硬脂酸2.85%,变异范围1.8%~3.9%;油酸46.36%,变异范围37.0%~64.7%;亚油酸32.86%,变异范围18.0%~40.4%;饱和脂肪酸含量19.21%,变异范围15.2%~22.1%。普通型花生的油酸含量高于其他类型,而亚油酸和棕榈酸含量低于其他类型。发掘出高油酸种质4份,低棕榈酸种质19份,低饱和脂肪酸种质7份。通过脂肪酸组成的分析,高油酸种质和低饱和脂肪酸种质均同时具备低棕榈酸的优良特性。SSR分析结果表明,这些种质的遗传差异相对较大。根据5对SSR引物的扩增结果,绘制了20份资源的分子指纹图谱,为这些优质资源的保护和有效利用奠定了基础。     

High-Oleic Peanut Oils Produced by HpRNA-Mediated Gene Silencing of Oleate Desaturase

The quality of peanut oil largely depends on the quantity of oleic (18:1) and linoleic acids (18:2). These two acids comprise more than 80% of the total fatty acids in peanuts. The oleate desaturase (FAD2) gene is important for maintaining high oleic acid content. A partial conservative sequence of the FAD2 gene from peanut was selected. The sense and antisense 260-bp fragments were amplified and subcloned into pFGC1008 binary expression vectors. A total of 21 transgenic plants were obtained via Agrobacterium-mediated transformation. The resulting down-regulation of the FAD2 gene resulted in a 70% increase in oleic acid content in the seeds of transformed plants compared with a 37.93% increase in untransformed plants. The results demonstrated that the target genes were likely suppressed by hpRNA interference, a pathway capable of achieving phenotypic changes. The silencing of FAD2 enabled the development of peanut oils having novel combinations of oleic acid content that can be used in high-value applications, making this approach a reliable technique for the genetic modification of seed quality and the potential for enhancement of other traits as well.     

Neuronal differentiation is triggered by oleic acid synthesized and released by astrocytes

Unlike in the adult brain, the newborn brain specifically takes up serum albumin during the postnatal period, coinciding with the stage of maximal brain development. Here we report that albumin stimulates oleic acid synthesis by astrocytes from the main metabolic substrates available during brain development. Oleic acid released by astrocytes is used by neurons for the synthesis of phospholipids and is specifically incorporated into growth cones. Oleic acid promotes axonal growth, neuronal clustering, and expression of the axonal growth-associated protein-43, GAP-43; all these observations indicating neuronal differentiation. The effect of oleic acid on GAP-43 synthesis is brought about by the activation of protein kinase C, since it was prevented by inhibitors of this kinase, such as H-7, polymyxin or sphingosine. The expression of GAP-43 was significantly increased in neurons co-cultured with astrocytes by the presence of albumin indicating that neuronal differentiation takes place in the presence of oleic acid synthesized and released by astrocytes in situ. In conclusion, during brain development the presence of albumin could play an important role by triggering the synthesis and release of oleic acid by astrocytes, which induces neuronal differentiation.     

Systematic study on ROS production induced by oleic, linoleic, and gamma-linolenic acids in human and rat neutrophils

The effects of oleic, linoleic, and gamma-linolenic acids on the production of ROS by unstimulated and PMA-stimulated neutrophils were investigated by using five techniques: luminol- and lucigenin-amplified chemiluminescence, cytochrome c, hydroethidine, and phenol red reduction. Using lucigenin-amplified chemiluminescence, an increase in extracellular superoxide levels was observed by the treatment of neutrophils with the fatty acids. There was also an increase in intracellular ROS levels under similar conditions as measured by the hydroethidine technique. An increment in the intra- and extracellular levels of H2O2 was also observed in neutrophils treated with oleic acid as measured by phenol red reduction assay. In the luminol technique, peroxidase activity is required in the reaction of luminol with ROS for light generation. Oleic, linoleic, and gamma-linolenic acids inhibited the myeloperoxidase activity in stimulated neutrophils. So, these fatty acids jeopardize the results of ROS content measured by this technique. Oleic, linoleic, and gamma-linolenic acids per se led to cytochrome c reduction and so this method also cannot be used to measure ROS production induced by fatty acids. Oleic, linoleic, and gamma-linolenic acids do stimulate ROS production by neutrophils; however, measurements using the luminol-amplified chemiluminescence and cytochrome c reduction techniques require further analysis.     

High Metastaticgastric and Breast Cancer Cells Consume Oleic Acid in an AMPK Dependent Manner

Gastric cancer and breast cancer have a clear tendency toward metastasis and invasion to the microenvironment predominantly composed of adipocytes. Oleic acid is an abundant monounsaturated fatty acid that releases from adipocytes and impinges on different energy metabolism responses. The effect and underlying mechanisms of oleic acid on highly metastatic cancer cells are not completely understood. We reported that AMP-activated protein kinase (AMPK) was obviously activated in highly aggressive carcinoma cell lines treated by oleic acid, including gastric carcinoma HGC-27 and breast carcinoma MDA-MB-231 cell lines. AMPK enhanced the rates of fatty acid oxidation and ATP production and thus significantly promoted cancer growth and migration under serum deprivation. Inactivation of AMPK attenuated these activities of oleic acid. Oleic acid inhibited cancer cell growth and survival in low metastatic carcinoma cells, such as gastric carcinoma SGC7901 and breast carcinoma MCF-7 cell lines. Pharmacological activation of AMPK rescued the cell viability by maintained ATP levels by increasing fatty acid β-oxidation. These results indicate that highly metastatic carcinoma cells could consume oleic acid to maintain malignancy in an AMPK-dependent manner. Our findings demonstrate the important contribution of fatty acid oxidation to cancer cell function.     

Oleic acid metabolism: A biochemical marker for studying photoregulation of epicotyl elongation in Vigna unguiculata

Oleic acid metabolism can be considered to be an indicator of growth photoregulation in cowpea ( Vigna unguiculata Westphal cv. M53) epicotyls. Phosphatidylcholine and phosphatidylethanolamine are the two lipid classes concerned in the photoregulation of oleic acid accumulation. The incorporation of radioactive precursors in internodes of whole plants has shown that there is de novo synthesis of these phospholipids during the light dependent growth process.
The variations in oleic acid content were used to study the photocontrol of elongation in segments excised from the apical part of the epicotyl. In this system, as in whole plants, oleic acid was the only fatty acid showing significant variation related to the light/dark treatments. Differences in photoresponse between excised intenode segments and internodes in whole plants are discussed.     

脂肪酶发酵过程中的油脂利用

对假丝酵母Candidasp．99—125发酵生产脂肪酶的过程中油脂代谢情况进行了研究。分析了发酵过程中甘油酯、油酸、棕榈酸等物质浓度随发酵时间的变化趋势,以及它们与菌体生长和产酶之间相互影响关系,结果发现油酸的消耗能够显著地促进脂肪酶的合成（油酸质量浓度从30g／L降低到10g／L）,并且细胞对油酸和棕榈酸的利用没有选择性,最终发酵脂肪酶活力可达8000U／mL。