胡须鸡β-防御素Ga1-1－Ga1-13基因克隆、序列分析及其在组织中分布术

用设计的特异性引物,采用RT-PCR技术扩增到胡须鸡β-防御素基因Gallinacin-1（Ga1-1）-Gallinacin-13（Ga1-13）共13个基因编码区全长片段。通过克隆、测序获得13个基因的cDNA核苷酸序列,GenBank登陆号为：DQ858311-DQ858323。比较分析胡须鸡13种β-防御素Ga1-1（a）-Ga1-13,Ga1-1基因与GenBank中注册的防御素基因氨基酸序列的同源性,均在96．5％-100％之间。利用Clustax软件对所获得的13种胡须鸡β-防御素基因进行系统发育树分析,结果显示Ga1-1、Ga1-1（a）、Ga1-2、Ga1-3、Ga1-4、Ga1-5、Ga1-6、Ga1-7、Ga1-8、Ga1-12和Ga1-13在同一大的分支上;Ga1-9和Ga1-10在同一分支;Ga1-11独在一分支上。用RT-PCR方法分析胡须鸡β-防御素Ga1-1－Ga1-13基因在不同组织中的分布,结果发现：Ga1-1、Ga1-2、Ga1-4、Ga1-5、Ga1-6、Ga1-7和Ga1-10的表达分布非常广泛,Ga1-3、Ga1-8、Ga1-9、Ga1-11、Ga1-12和Ga1-13的分布少。     

2021年清远市香稻新品种区域试验

为筛选出适合广东省清远市种植的高产优质香稻新品种,为水稻新品种审定、推广及品种结构调整提供科学依据,2021年开展了10个香稻新品种的区域试验.初步结果表明,又美优219、又美优珍香、众香5号和粤品优珍香4个品种的综合表现优良,建议进一步扩大区域试验.     

生态条件对大麦产量及β-葡聚糖含量的影响

为明确不同生态条件下大麦籽粒产量及β-葡聚糖含量的环境效应,选取了11个不同的大麦品种,在全国6个生态条件有差异的试点种植。结果表明,各试点间大麦籽粒产量差异显著,襄樊试点平均产量最高;苏啤3号大麦在6个试点的平均产量最高,达5391.7kg/hm2,与其他品种差异显著。各试点及各品种间,大麦籽粒β-葡聚糖含量差异也达到显著水平。G231M004M大麦在6个试点β-葡聚糖含量的平均值显著高于其他品种;各个大麦品种在保山试点的籽粒β-葡聚糖含量均明显高于其他试点。因此,选取特定品种种植在适宜的生态条件下对调节大麦籽粒产量和β-葡聚糖含量具有积极意义。     

利用CRISPR/Cas9创造早熟香味水稻

黑龙江省是我国最大的水稻种植区,近年来水稻种植面积不断增加,为加快高纬度、低积温的地区品种选育进程,本研究利用CRISPR/Cas9基因编辑技术对第二积温带主栽品种绥粳14的抽穗期基因Hd2、香味基因Badh2进行编辑。通过T_1代的分子鉴定,Hd2和Badh2基因被敲除而且基因型纯合。田间鉴定表明,转基因抗性标记也被成功剔除的两个株系,绥粳14-d1和绥粳14-d2,比绥粳14提早13天左右。利用咀嚼法、氢氧化钾浸泡法和气相色谱-质谱联用技术(GCMS)测定香味,结果显示绥粳14-d1和绥粳14-d2中香味物质2-乙酰-1-吡咯啉(2-AP)含量显著高于绥粳14。综合以上结果,本研究利用基因编辑技术,成功获得了改良的抽穗期缩短香米品种,为早熟、优质水稻品种的选育提供了理论指导,加速了育种进程。     

紫苏新品系2012-8-1选育报告

紫苏新品系陇苏2012-8-1是甘肃省农业科学院生物技术研究所在对紫苏种质鉴定及品种优选的基础上,利用多代集团混合选择技术,经过连续5 a选育而成。2016 — 2017年参加甘肃省多点区域试验,2 a 10点(次)平均折合产量为3 117.15 kg/hm2,较对照品种陇苏1号增产18.57%。该品系生育期158 d,平均株高168.1 cm,籽粒灰褐色,千粒重3.59 g,单株产量74.81 g。田间表现丰产性、稳产性好,抗病性强。籽粒α-亚麻酸含量58.35%,含油44.66%,饱和脂肪酸含量19.45%,不饱和脂肪酸含量78.56%,品质优良。适宜在甘肃省庆阳、平凉、天水等半干旱山区及其同类地区种植。     

桂花15-顺式-ζ-胡萝卜素异构酶基因(Z-ISO)的克隆及表达分析

15-顺式-ζ-胡萝卜素异构酶(15-cis-ζ-carotene isomerase,Z-ISO)可以催化9,15,9'-三顺式-ζ-胡萝卜素异构化生成9,9'-二顺式-ζ-胡萝卜素,Z-ISO基因是植物类胡萝卜素代谢途径中最后一个被鉴定的基因.为克隆桂花(Osmanthus fragrans) Z-ISO基因的ORF序列并研究其表达特征,本研究利用已构建的桂花花瓣转录组数据库中Unigene序列信息,结合PCR技术克隆得到桂花Of Z-ISO1(GenBank登录号:KX120175)和Of Z-ISO2(GenBank登录号:KX120176)基因ORF序列.生物信息学分析表明,Of Z-ISO1和OfZ-ISO2基因均含有长为1 107 bp的ORF,编码368个氨基酸残基.Of Z-ISO1和Of Z-ISO2蛋白均含有6个跨膜螺旋区和3个保守的异构酶功能作用所必需的活性位点(H152、H268和D296).Of Z-ISO1和Of-ISO2与玉米(Zea mays)和拟南芥(Arabidopsis thaliana) Z-ISO1氨基酸序列有较高的相似性,系统进化树分析发现,Of Z-ISO1和Of Z-ISO2与芝麻(Sesamum indicum)Z-ISO亲缘关系最近,聚为同一小支.qRT-PCR结果显示,堰虹桂花序发育过程中,OfZ-ISO1基因在花蕾期花序中表达量较低,铃梗期其表达量急剧增加,初开期表达量保持不变,而盛开期表达量显著下降;Of Z-ISO2基因在花蕾期和铃梗期花序中表达量较低,随后表达量逐渐增加,于盛开期表达量达到最高.在桂花不同花色品种中,丹桂品种堰虹桂花色最深、花瓣类胡萝卜、素总量最高,金桂品种金球桂次之,银桂品种玉玲珑花色最浅、花瓣类胡萝卜素总量最低;总体上铃梗期和初开期时玉玲珑Of Z-ISO1和Of Z-ISO2基因的表达量较高,盛开期3个不同花色品种花瓣中OfZ-ISO1和Ofz-ISO2基因的表达量大致相同.本研究结果表明,桂花花序开放过程中Of Z-ISO1和Of Z-ISO2基因的表达水平对其类胡萝卜素的积累十分重要,不同花色品种类胡萝卜素含量与其Of-ISO1和OfZ-ISO2基因表达量并非正相关.研究结果将为全面揭示桂花呈色机制提供理论基础.     

香稻产量和品质对化肥施用及气候因素的响应

  【目的】  香稻因其独特的香味深受消费者喜爱,明确香稻产量和品质对化肥施用的响应及其与气候条件的关系,对综合提升香稻产量和品质具有重要意义。  【方法】  在中国知网、谷歌学术等数据库以关键词“水稻”、“香气”、“香味”以及“香稻”和“香米”及其组合进行文献检索,共获得501组试验数据对。将施肥与对照分为两组,采用整合分析方法(meta-analysis),分析了施用氮磷钾肥对香稻产量和品质的影响,以及在不同生育期累积降水量、积温和日照时数条件下的变化率。  【结果】  施用氮、磷、钾肥,香稻的平均增产率分别为23.7%、25.8%、18.0%。与不施用氮肥相比,施用氮肥对粳稻和籼稻增产率分别为27.6%和20.2%;与不施用钾肥相比,施用钾肥对粳稻和籼稻的增产率分别为29.9%和7.3%。施用氮肥可显著增加香稻碾磨品质(整精米率)和蒸煮品质(粗蛋白含量,胶稠度),施用钾肥可显著增加香稻碾磨品质(整精米率)、蒸煮品质(粗蛋白含量,胶稠度)、外观品质(垩白度)和香味品质(2-乙酰-1-吡咯啉含量,以下简称2-AP)。施用氮肥对籼型常规香稻的品质提升效果显著,而施用钾肥则对籼型杂交香稻的品质提升更显著。此外,施用氮肥下香稻整精米率的变化率随生育期内有效积温升高呈持续增加趋势,而随累积降水量和日照时数的增加呈先升后降趋势;胶稠度的变化率随气候变化(有效积温、降水量和日照时数等的增加)呈现先降后增趋势。施用钾肥下香稻整精米率的变化率随生育期内有效积温和日照时数升高呈现先降后增趋势,而随累积降水量的变化呈先升后降趋势;垩白粒率的变化率随气候变化均呈先降后增的趋势;胶稠度的变化率随气候变化(有效积温、降水量和日照时数等的增加)均呈降低的趋势。  【结论】  施用氮肥或钾肥均有助于香稻产量和品质的协同提升,且对籼型香稻增产提质效果更显著。同时生育期内累积降水量、有效积温、日照时数影响着氮肥和钾肥的产量和品质效应,分别达到800～1100 mm、2900～3400 ℃·d和800～1000 h时,结合氮肥调控可协同提高香稻产量、加工品质和蒸煮品质;分别达到500～800 mm、2900～3400 ℃·d和400～600 h时,结合钾肥调控可协同提高香稻产量、加工品质、蒸煮品质和外观品质。     

静宁县谷子新品种(系)引种试验初报

以陇谷11号为对照,在静宁县对7个谷子新品种(系)进行了引种试验。结果表明,在露地栽培条件下,籽粒产量以9410-4-2-2-1最高,达4 768.52 kg/hm2,较对照品种陇谷11号增产22.91%;0416-2-1-1-1、0412-1-2-1、029-5-5-3分别较对照增产22.20%、11.22%、4.30%,且综合表现优良。以上品种(系)适宜在静宁县山旱梯田地露地种植。     

胡须鸡β-防御素Gal-1-Gal-13基因克隆、序列分析及其在组织中分布

用设计的特异性引物,采用RT-PCR技术扩增到胡须鸡β-防御素基因Gallinacin-1(Gal-1)～Gallinacin-13(Gal-13)共13个基因编码区全长片段.通过克隆、测序获得13个基因的cDNA核苷酸序列,GenBank登陆号为:DQ858311～DQ858323.比较分析胡须鸡13种β-防御素Gal-1(a)～Gal-13,Gal-1基因与GenBank中注册的防御素基因氨基酸序列的同源性.均在96.5%～100%之间.利用Clustax软件对所获得的13种胡须鸡β-防御素基因进行系统发育树分析,结果显示Gal-1、Gal-1(a)、Gal-2、Gal-3、Gal-4、Gal-5、Gal-6、Gal-7、Gal-8、Gal-12和Gal-13在同一大的分支上;Gal-9和Gal-10在同一分支;Gal-11独在一分支上.用RT-PCR方法分析胡须鸡β-防御素Gal-1～Gal-13基因在不同组织中的分布,结果发现:Gal-1、Gal-2、Gal-4、Gal-5、Gal-6、Gal-7和Gal-10的表达分布非常广泛,Gal-3、Gal-8、Gal-9、Gal-11、Gal-12和Gal-13的分布少.     

利用CAPS标记推测花生品种(系)FAD2位点基因型的研究

以5个高油酸含量花生品种(系)和2个普通油酸含量花生品种为试材,利用CAPS标记对这些品种(系)的FAD2位点基因型进行分析.结果表明:5个高油酸含量花生品种(系)的油酸含量均在80％以上,普通油酸含量品种花育22号和花育28号油酸含量分别为51.62％和41.38％.基于CAPS标记在7个品种(系)中的扩增产物及酶切带型分析推测,AhFAD2A位点花育28号为野生型,其他6个品种(系)符合448 G＞A突变类型;AhFAD2B位点花育28号和花育22号为野生型,开农H03-3、花育32号、P76和F18符合441-442insA突变类型,而06B16在该位点不符合441-442insA突变类型.     

Biosynthetic mechanism of 2-acetyl-1-pyrroline and its relationship with Delta1-pyrroline-5-carboxylic acid and methylglyoxal in aromatic rice (Oryza sativa L.) callus

2-Acetyl-1-pyrroline (2-AP) was identified as the major flavor compound in aromatic rice varieties Tainung 71 and 72. In order to understand the mechanism of 2-AP biosynthesis in aromatic rice, we studied the formation of putative precursors, Delta(1)-pyrroline-5-carboxylic acid and methylglyoxal. The endogenous Delta(1)-pyrroline-5-carboxylic acid contents of Tainung 71 and 72 calli reached 191 to 276%, compared to nonaromatic rice Tainung 67. In addition, calli of Tainung 71 and 72 contained 1.30- and 1.36-fold, respectively, higher methylglyoxal levels than that of Tainung 67. Specific enzyme activities of Delta(1)-pyrroline-5-carboxylic acid-synthetic enzyme including Delta(1)-pyrolline-5-carboxylic acid synthetase (P5CS) and ornithine aminotransferase (OAT) increased significantly in aromatic rice varieties. The expression levels of P5CS1 and P5CS2 genes were found to be significantly higher in aromatic rice than nonaromatic rice. Results of a tracer experiment with (15)N-labeled glutamic acid revealed that the nitrogen atom of 2-acetyl-1-pyrroline was derived from glutamic acid. Upregulation of P5CS in aromatic rice Tainung 72 may contribute to the increase of Delta(1)-pyrroline-5-carboxylic acid level and thus leads to the accumulation of an extra amount of 2-acetyl-1-pyrroline.     

Screening for 2-acetyl-1-pyrroline in the headspace of rice using SPME/GC-MS

Solid phase microextraction (SPME) is used to collect and concentrate the compounds in the headspace of rice. This research describes optimization parameters of temperature, moisture, and sampling time. Optimization was based upon the recovered levels of 2-acetyl-1-pyrroline (2-AP), the popcorn aroma in aromatic rice. The method uses a sampling temperature of 80 degrees C and adds 100 microL of water to a 0.75 g sample of rice. The rice was preheated for 25 min, a carboxen/DVB/PDMS SPME fiber was exposed to the headspace for 15 min, and a subsequent GC-MS analysis took 35 min. Samples of rice can be analyzed as the flour, milled kernels, or brown rice. Twenty-one experimental rice varieties were analyzed by the SPME method and compared to a wet technique. Recoveries of several nanograms of 2-AP from 0.75 g samples of aromatic rice were observed, whereas only trace amounts of 2-AP were recovered from nonaromatic rice. Recovery from a single SPME headspace analysis is calculated to be 0.3% of the total 2-AP in the sample.     

Precursors of 2-acetyl-1-pyrroline, a potent flavor compound of an aromatic rice variety

The biological formation of a potent flavor compound, 2-acetyl-1-pyrroline, in the aromatic rice variety (Khao Dawk Mali 105) was studied in seedlings and callus of the rice. Concentrations of 2-acetyl-1-pyrroline were determined by GC-MS-SIM using an isotope dilution method. Increases in concentration occurred when proline, ornithine, and glutamate were present in solution, with proline increasing the concentration by more than 3-fold compared to that of the control. Results of tracer experiments using (15)N-proline, (15)N-glycine, and proline-1-(13)C indicated that the nitrogen source of 2-acetyl-1-pyrroline was proline, whereas the carbon source of the acetyl group was not the carboxyl group of proline. 2-acetyl-1-pyrroline was formed in the aromatic rice at temperatures below that of thermal generation in bread baking, and formed in the aerial part of aromatic rice from proline as the nitrogen precursor.     

Aroma of Cooked Rice (Oryza sativa): Comparison Between Commercial Basmati and Italian Line B5-3

Selected lines of aromatic rice from American cv. A301 were cultivated in Italy, and the agronomic traits and chemical properties related to their aroma were studied. The most characteristic compound responsible for aromatic rice, 2-acetyl-1-pyrroline, was quantified in all lines. Line B5-3, characterized by high 2-acetyl-1-pyrroline content, was investigated in greater detail for its volatile components and was compared with a commercial Basmati rice. Volatiles were collected by steam-distillation. Several classes of compounds were identified and quantified in both samples, including hydrocarbons, aldehydes, alcohols, ketones, heterocyclic, terpenes, disulfides, and phenols. Hexanal was the most abundant compound in both samples, followed by pentanal, 2-acetyl-1-pyrroline, hexanol, benzaldehyde, oct-1-en-3-ol, 4-vinylguaiacol, indole, and trans-2- nonenal. 2-Acetyl-1-pyrroline was present at 570 and 2,350 ppb in Basmati and B5-3, respectively.     

Distribution of total aflatoxins in milled fractions of hulled rice

Two varieties of hulled rice artificially contaminated with aflatoxins at five different levels were processed by dehulling and polishing methods. Contamination levels ranged from 356 to 818 microg/kg and from 244 to 645 microg/kg in medium and long grain rice, respectively. After physical processing, four different milled fractions were obtained (hull, bran, polished broken grains, and polished whole kernels). The fractions were analyzed for total aflatoxins (B1, B2, G1, and G2) by enzyme-linked immunosorbent assay (ELISA). Aflatoxins were removed in fractions intended for human consumption (polished broken grains and polished whole kernels) at rates up to 97%. They were found throughout all fractions, but higher contamination levels were detected in hull and bran fractions than in unprocessed kernels and polished fractions. Regardless of the rice variety, the aflatoxin distribution pattern depended on the initial contamination level and type of milled fraction but not on the duration of polishing.     

Identification and quantitation of the rice aroma compound, 2-acetyl-1-pyrroline,in bread flowers (Vallaris glabra Ktze)

The aroma impact compound, 2-acetyl-1-pyrroline (2AP), has been identified for the first time in headspace of fresh bread flowers (Vallaris glabra Ktze) in which volatile components were extracted by solid-phase microextraction (SPME) at room temperature prior to analysis by gas chromatography-mass spectrometry. A total of 50 volatiles were detected. Among these, 23 volatiles were identified, predominantly in a group of terpenes. More volatiles were found in the extract of fresh bread flowers obtained by continuous steam distillation and solvent extraction (SDS). Of the 40 volatiles identified, the additional components were mainly straight-chain saturated hydrocarbons. 2AP was found in the extracts obtained by both SPME (0.37%) and SDS (2.71% relative proportion). Quantitative analyses of 2AP in bread flowers and other plant materials were performed by solvent extraction employing acidic solutions and capillary GC with flame ionization detection. The highest concentration of 2AP was found in dried flowers of V. glabra at 26.1 mg/kg. By comparison with other plant sources, fresh leaves of Pandanus amaryllifolius Roxb contain 2AP at 10.3 mg/kg and Thai fragrant rice, Khao Dawk Mali 105, at 3.0 mg/kg.     

南方6省稻米总汞含量调查及其膳食暴露评估

为评价中国南方6省稻米中总汞污染现状,于2009年在江西、湖北、湖南、广东、广西和四川6省抽样检测了1 321份稻米样品,结合中国居民20个性别年龄组人群的稻米消费量和体重信息,采用非参数概率方法对稻米中总汞的膳食暴露量进行了评估。总汞含量采用电感耦合等离子体质谱仪（ICP-MS）测定,检出限（LOD）为0.000 8 mg.kg-1。结果表明,稻米的总汞含量在地区间存在着差异,虽然有76.2%的样本总汞含量（0.000 8～0.063 4 mg.kg-1）高于检出限,但仅有2.3%的样本超出了最高限量（ML 0.02mg.kg-1）。将评估结果比照JECFA推荐的总汞暂定每周耐受摄入量（PTWI）5μg.（kg bw）-1,我国居民食用这6省稻米产生的汞暴露风险较小。但在P99.9的高百分位水平下,14岁以下人群的摄入量相对较高,占PTWI的41.5%～62.9%,其中2～4岁儿童和4～7岁男童的摄入量占PTWI的60%以上,潜在风险较大。建议应对稻米中重金属汞的含量进行追踪监测。     

氮锌肥配施对南方香粳稻产量和品质的影响

【目的】研究氮锌肥配施对南方香粳稻产量和外观加工品质、蒸煮食味品质、香气的影响,为南方香粳稻的调优栽培提供科学依据。【方法】田间试验于2020和2021年在扬州大学试验基地进行,供试粳稻品种为宁香粳9号和南粳46。在施锌(+Zn)和不施锌(Zn0)条件下,分别设3个施氮水平：0 (N0)、180 kg/hm²(N180)、270 kg/hm²(N270),共6个处理。于收获期,测定产量和产量构成因素,分析籽粒加工外观、食味和香气品质。【结果】提高氮肥用量可显著增加香粳稻产量,而施用锌肥对香粳稻产量无显著影响,+ZnN270处理宁香粳9号和南粳46产量均最高,分别达10.48 t/hm²和10.40 t/hm²;相同氮肥用量下,增施锌肥对产量没有显著影响。宁香粳9号和南粳46的最佳稻米加工和外观品质均以+ZnN270处理最优。在相同氮肥用量下,施用锌肥改善了香粳稻的加工和外观品质。与N0处理相比,在N180和N270处理下两品种的直链淀粉含量下降了8.44%～13.16%,蛋白质含量增加了9.79%～...     

Quantitation of tocotrienol and tocopherol in various rice brans

Rice bran is abundant in bioactive compounds including tocotrienol (T3, unsaturated vitamin E). T3 has been reported about its potential functionalities (i.e., antiangiogenic effect), so much attention has been paid on usability of rice bran T3. Hence, we developed a rapid screening method for T3-rich rice bran by one-step equilibrium direct solvent extraction followed by normal phase high-performance liquid chromatography (HPLC). The method gave high-extraction rate of rice bran T3 and tocopherol (above 90%), and the determination of vitamin E by HPLC was completed within 15 min. Using the method, an average of total T3 content in 109 kinds of rice bran samples was 830 mug/g dry wt. Kouchi-Akamai, Joushuu, and Wataribune were found as the T3-rich rice bran varieties (1350-1430 microg T3/g dry wt). According to T3 ratio against total vitamin E (wt %), the average ratio was 61%. Hirayama, Moritawase, and Kaneko were found as the varieties having the highest T3 ratio (80-86%). Since T3 content in Koshihikari rice bran (the leading variety in Japan) was a little above the average, we cross-fertilized Koshihikari with T3-rich varieties and found that T3 content or ratio in F1 was improved compared with Koshihikari. The varieties found rich in T3 could be used for nutraceutical purpose.     

Nutraceutical Concentrations within the Bran of Various Rice Kernel Thickness Fractions

Several important nutraceutical compounds, such as tocotrienols, tocopherols, and oryzanols, can be extracted from rice bran, a by-product of milling. This study was conducted to not only provide information regarding nutraceutical concentrations within the rice kernel based on bran collected from successive milling, but also to determine levels of nutraceutical concentrations across several different thickness fractions. Nutraceutical compounds were measured in the bran from two long-grain rice varieties, Cypress and Drew. Rough rice was separated into three thickness fractions (<1·84, 1·84–1·98, and >1·98 mm) and each fraction milled for three successive 10 s milling durations. Bran was collected from each milling duration of each thickness fraction to allow quantification of the nutraceutical content. Results showed that bran collected from rice milled for longer durations (30 s) had lower levels of tocotrienols and tocopherols compared to bran from shorter milling durations (10 s). The highest concentration of oryzanols was in the rice bran from the first 10 s milling duration. Overall, compared to bran from thinner kernels (<1·84 mm), the bran from thicker kernel fractions contained a higher content of nutraceuticals.