西藏南美藜（Chenopodium quinoa Willd）病害初步研究

本文研究报道１９９３～１９９４年南美黎在西藏病害的发生情况，经鉴定有真菌病害７种，病毒病害１种。并提出防治建议。

Developing Genetic Variability of Quinoa (<i>Chenopodium quinoa</i>Willd.) with Gamma Radiation for Use in Breeding Programs

Quinoa (Chenopodium quinoa Willd.) is a staple food produced mainly by small-scale subsistence farmers in Peru’s highland. Dry seeds (cv. Pasankalla) were irradiated with doses of 150 Gy, 250 Gy and 350 Gy. In the M1 generation, the germination process was delayed with increasing radiation dose;seedling height, root length and leaf development were most reduced at 250 Gy and at 350 Gy, no plants survived. In M2, the maximum spectrum of chlorophyll mutations corresponded to 150 Gy and the maximum frequency to 250 Gy. The chlorine mutation was predominant, followed by xantha. Changes were registered for branch number, pedicel length, plant height, life-cycle duration, stem and foliage colour, and leaf morphology at the two doses, with improvements in plant type. More than one mutation per plant was found, especially at 250 Gy. In M3, the same spectrum of mutations was observed, along with a valuable change in grain colour.

Present Situations and Suggestions for Large-scale Development of Chenopodium quinoa Willd.

In this paper,based on the study of Chenopodium quinoa Willd.planting,it is concluded that the current situations of large-scale development of C.quinoa Willd.include the following four aspects:first,the research on C.quinoa Willd.varieties needs to be strengthened;second,it is insufficient to master the training environment and cultivation techniques;third,the large-scale planting of C.quinoa Willd.is not enough;fourth,the degree of mechanization of C.quinoa Willd.planting is not enough.In view of the above situation,this paper puts forward the following effective suggestions to strengthen the large-scale development of C.quinoa Willd.:the first is to increase the investment in C.quinoa Willd.variety research;the second is to strengthen the analysis of introduction and screening of C.quinoa Willd.varieties;the third is to study the best planting environment and cultivation techniques of C.quinoa Willd.;the fourth is to increase the scale of mechanized production of C.quinoa Willd.;the fifth is to increase the research and development of C.quinoa Willd.related products and their deep processing technology.

Rapid Detection of Fat Content in Chenopodium quinoa Willd by Near Infrared Spectroscopy

This study was conducted to find a method for rapid determination of fat content in complete quinoa （ Chenopodium quinoa Willd） seeds. The near infrared spectra of 100 quinoa samples were collected, and a mathematic model was built using the near infrared spectra, so as to perform prediction. The results showed that within the wavelength range of 1 0 000-4 000 cm ^-1 , the quantification model of fat content built by first derivative ＋vector normalization spectral pre-processing had better calibration and prediction effects, and showed a determination coefficient of cross validation （ r cv^ 2 ） of 0.939 3 and a determination coefficient of validation （ rval^2 ） of 0.923 5. The near infrared spectral model of fat could be used for rapid detection of fat contents in quinoa.

Rapid Detection of Starch Content in Quinoa(Chenopodium quinoa Willd.)by Near Infrared Spectroscopy

This study was conducted to establish a method for rapid determination of crude starch content in complete quinoa （ Chenopodium quinoa Willd） seeds. The near infrared spectra of 100 quinoa samples were collected, and a mathematic model was built using the near infrared spectra within the wavelength range of 1 0 000-4 000 cm^-1 by first derivative ＋vector normalization spectral pre-processing. The results showed that the quantification model of starch content had better calibration and prediction effects, and showed a determination coefficient of cross validation （ r^2 cv ） of 0.914 7 and a determination coefficient of validation （ r^2 val ） of 0.903 1. The determination of starch content in complete quinoa seeds by near infrared spectroscopy is totally feasible.

藜麦(Chenopodium quinoa)内生菌LMJB-14的鉴定与生防促生功能研究

采用平板对峙法,从藜麦内生菌中筛选出对链格孢属(Alternaria sp.)、镰孢菌属(Fusarium sp.)、刺盘孢属(Colletotrichum sp.)3种常见植物真菌性病害具有较强拮抗作用的细菌,通过菌悬液浸种实验测定其对藜麦种子的促生效果,结合形态学特征、生理生化特征、基因测序分析以确定其分类学地位。结果表明:细菌LMJB-14对3种常见植物病原真菌具有不同程度的拮抗效果,其中对刺盘孢属病原菌拮抗效果最好(抑菌率为31.2%),其次为镰孢菌属病原菌(抑菌率为23.2%),对链格铇属病原菌抑菌率最低(19.2%);以菌株LMJB-14菌悬液浸种对藜麦种子萌发具有促进作用;经鉴定该拮抗菌株为贝莱斯芽孢杆菌。

Evolutionary divergence in Chenopodium and validation of SNPs in chloroplast rbcL and matK genes by allele-specific PCR for development of Chenopodium quinoa-specific markers

The genus Chenopodium comprises about 150 species, of which Chenopodium quinoa and C. album are important for their nutritional value. Evaluation of variation in qualitative morphological traits of plants and SNPs in chloroplast rbc L and mat K gene sequences in 19 accessions representing C. quinoa and C. album indicated that the accessions IC-411824 and IC-411825,which have white seeds, belong to C. quinoa rather than C. album. This observation was also supported by a time tree that indicated IC-411824 and IC-411825 to be a sister clade to accessions of C. quinoa with an estimated age of 1.2 Mya. Whereas multiple alignments of rbc L gene sequences from the 19 accessions revealed 1.26% parsimony-informative sites with 0.68%interspecific sequence diversity, alignment of nucleotide sequences of amplicons representing the mat K gene revealed 4.97% parsimony-informative sites and 2.81% interspecific sequence diversity. Validation of SNPs in the cp rbc L and mat K regions of 36 accessions belonging to C. quinoa and C. album was performed by allele-specific PCR with primers carrying a single base change at the 3′ end. We report the first C. quinoa-specific SNP-based primer, R1RQ-AFR,designed from rbc L sequences, that could differentiate quinoa from 64 genera including13 species of the genus Chenopodium. With an estimated age of 10.5–4.1 million years(Myr), the Himalayan chenopods are evolutionarily younger than the Andean chenopods. The results establish the paraphyletic origin of the genus Chenopodium.

CHENOPODIUM QUINOA:Super -Nutritious Food for the 21st Century

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未来超级营养作物—昆诺阿藜(QUINOA)

昆诺阿藜是南美安底斯高原古老的作物;至少已有五十多年的种植历史.印加人称之为“谷物之母”。和其它作物相比,昆诺阿藜会有更高的蛋白质、钙质、铁质,有些研究者认为:还没有一种食物能象它那样提供全面的人体所需的基本营养,无论在植物界或动物界.昆诺阿藜在分类学上属于藜科(Chenopodiaceue),学名为 Chenopodium qu-inoa,其叶形等许多特征都很象通常我们所见的灰灰菜,其籽粒很小呈上下平坦的圆盘状,浅黄色、白色或粉红色,但有些品种呈红色,橙黄色或黑色。

Agronomic and Productivity Performance for Quinoa Genotypes in an Agroecological and Conventional Production System

The objective of this research was to evaluate the agronomic performance and productivity of sixteen genotypes of Chenopodium quinoa cultivated in agroecological and conventional production systems. The evaluations were carried out, based on agronomic characteristics and yields of sixteen C. quinoa genotypes, grown in two simultaneous experiments, in an agroecological production and a conventional production system carried out at the town of Entre Rios do Oeste, Paraná, Brazil in the harvest 2015/16. Each experiment was composed of three replicates, following the randomized block design. Number of plants in flowering, number of plants per linear meter, height of insertion of the first panicle, number of days for maturation and productivity were the parameters evaluated. The data were submitted to statistical analysis with the aid of the GENES computational application. Genotype Q13-24 showed a more suitable production for the conventional production system. While the genotype Q13-01, presented the increase of productivity, being more indicated to the system of agroecological production. The characteristics height of plant flowers (HPF) and height of insertion of the first panicle (HIP) had higher values when the plants were cultivated in a conventional system. The number of plants per linear meter (NPLM) was higher in the agroecological crop, when compared to conventional cultivation. The same quinoa genotype can behave differently depending on the area management, being a productivity and the genotype cycle depends on the production system and the genotype used.

36份藜麦种质资源苗期耐盐碱性评价与筛选

为探究不同藜麦种质苗期耐盐碱性差异,本研究对36份藜麦种质苗期用150 mmol/L盐碱浓度(NaCl∶NaHCO_(3)=4∶1)进行胁迫处理,对株高、叶面积和叶片相对含水量等11个指标进行了测定,采用主成分分析等多种统计方法,筛选出适宜山西地区生长的藜麦种质。结果表明,盐碱胁迫下,藜麦种质各类指标相互之间均存在一定程度的关联;本试验中共提取5个主成分因子,方差累积贡献率为74.947%,可以代替11个指标的绝大部分信息,显示盐碱胁迫下藜麦苗期的抗盐碱特性,并且计算出耐盐碱性综合评价D值,并基于D值进行聚类分析,将36份种质资源分为四个大类,包括高耐盐碱种质5份、耐盐碱种质14份、弱耐盐种质11份、盐碱敏感种质共6份。

干旱胁迫对不同品系藜麦苗期生长特性的影响

为了给干旱地区的藜麦品种选择提供理论支持和技术指导,分析研究了不同浓度聚乙二醇(PEG-6000)对藜麦苗期生长特性的影响。结果表明,不同品系的藜麦在干旱胁迫下的生长和生理响应存在差异,随着干旱胁迫的增强,藜麦的株高、叶面积和生物量呈下降趋势;不同品系藜麦对干旱胁迫的响应差异表明了它们的抗旱特性的不同,黑色和红色品系的藜麦在一定胁迫程度下表现出较好的适应性和较高的抗旱能力,而灰色和白色品系的藜麦对干旱胁迫的敏感性相对较高。因此,在干旱地区进行藜麦种植时,选择具有较强抗旱能力的黑色和红色品系会更加适宜。

烟剂对大棚藜麦菜光合作用和叶绿素含量的影响

在藜麦菜的温室大棚种植中,用12%百菌清烟剂与12%速克灵烟剂防治病害较为常用。为了研究这两种烟剂对藜麦菜的叶绿素含量和光合作用的影响,施用烟剂后第3天和第5天分别进行藜麦菜叶片叶绿素含量和光合速率的检测。结果显示,施用烟剂可以提高藜麦叶绿素的含量,提高光合速率,进而提高生产率。

藜麦SOD家族基因的鉴定及其对混合盐碱胁迫的响应

超氧化物歧化酶(superoxide dismutase,SOD)是植物抗氧化系统的关键酶,在保护植物免受生物和非生物胁迫方面发挥重要作用。以拟南芥SOD为基础,通过序列比对在藜麦基因组中鉴定出12个SOD基因,分别定位于细胞核、微体及线粒体,在11条染色体上不均匀分布,其编码蛋白质三级结构显示Cu/Zn-SODs与Fe-SODs为同源二聚体,Mn-SODs为同源四聚体。CqSOD基因内含子/外显子分布模式不尽相同,内含子数介于4~7个,保守基序差异明显。系统发育关系显示,SOD蛋白可分为Cu/Zn-SODs、Fe-SODs及Mn-SODs 3个亚族。此外,所有的CqFe-SODs及CqMn-SODs启动子区都含有脱落酸激素反应顺式元件,CqSOD12与11个CqSOD蛋白及4个CqCAT蛋白存在相互作用。表达谱分析表明,12个CqSOD基因对混合盐碱及硝普钠均有较强响应。研究结果为SOD基因在植物发育和胁迫响应中的作用及分子机制研究奠定基础。

正交试验优化藜麦(Chenopodium quinoa)离体再生体系

为了优化藜麦离体再生体系的培养条件,实现藜麦离体高效快速繁殖。本试验选取材料‘忻藜1号’茎段和子叶为外植体,采用正交设计筛选出最适于藜麦愈伤组织分化的激素(6-苄氨基嘌呤,萘乙酸)、光强组合。结果显示:不同外植体在MS+6-BA 1.5 mg/L+NAA 0.4 mg/L培养基中出愈率最高,且茎段比子叶诱导率高,分别为77%和60%;在光强为4000 lx以上,MS+6-BA 1.5 mg/L+NAA 0.2 mg/L培养基中,不定芽分化率最高,可达96%。本试验的正交设计为藜麦快速繁殖和遗传转化提供技术支持。

藜麦CqGAI基因密码子偏好性与进化分析

为了阐明藜麦CqGAI基因密码子使用特性,克隆获得藜麦CqGAI基因序列,运用CodonW、SPSS软件及EMBOSS在线程序分析藜麦CqGAI基因密码子使用偏好性,并与25种植物的GAI基因进行中性绘图、ENC分析和奇偶偏好偏差性分析。结果表明,藜麦CqGAI基因编码序列(coding sequence,CDS)全长1 782 bp,编码593个氨基酸,包含DELLA基因家族特有结构域DELLA、TVHYNP、NLS、VHIID、LHR、RVER;CqGAI基因能够迅速响应赤霉素(gibberellins,GA),在GA信号通路中起关键作用;密码子偏好性分析显示,CqGAI基因的有效密码子数(effective number of codon,ENC)、密码子适应指数(codon adaptation index, CAI)及GC含量分别为54.14、0.21和46.18%,密码子偏好性较弱,偏好以A/T结尾,有27个高频密码子。聚类分析表明,藜麦CqGAI基因与石竹目植物的亲缘关系较近。碱基组成与相关性分析发现,CqGAI基因密码子偏好性受碱基突变和选择效应影响。密码子使用频率表明,大肠杆菌与酵母菌均适用于藜麦CqGAI基因异源表达,拟南芥、烟草、甜菜均可作为藜麦CqGAI基因功能分析的遗传转化受体。以上研究结果为进一步研究藜麦CqGAI基因功能和异源表达提供了重要参考。

31份藜麦种质农艺性状及ISSR遗传多样性分析

【目的】筛选西北干旱及半干旱地区不同利用价值藜麦种质,为藜麦种质资源的保护与利用奠定基础。【方法】以种植于甘肃临夏东乡县的31份藜麦种质资源为材料,分析其农艺性状,利用ISSR引物进行分子标记PCR扩增,阐明其遗传多样性及特点。【结果】株高、茎粗、鲜干比、茎叶比、单株产量5个农艺性状的变异系数在14.41%~63.30%,其中株高与茎粗、鲜干比呈极显著正相关,与单株产量呈显著正相关。茎粗与单株产量呈极显著正相关,与鲜干比呈显著正相关。鲜干比与单株产量呈极显著正相关。筛选出8条多态性高且清晰的ISSR引物,扩增出63条条带,其中多态性条带55条,多态性位点比率为83.70%,有效等位基因数(Ne)为1.4515,基因多样性(H)为0.2731,香农信息指数(I)为0.4174。根据UPGMA聚类分析结果,在遗传相似系数为0.72时,31份藜麦种质可分为5个类群。第1类群为已选育的陇藜1号、陇藜7号,抗倒伏性、抗病虫害好;第2类群株高较低,可作为抗倒伏材料进一步选育;第3类群株高高、单株产量大、鲜干比适中、茎叶比小,可作为鲜喂材料进一步选育;第4类群鲜干比最大,且与台湾红藜LQ-223的亲缘关系相近;第5类群具有茎叶比最大、中熟品种的特性。【结论】31份藜麦种质资源具有较高的遗传多样性,可为藜麦种质资源创新、遗传育种提供优质材料和科学依据。

藜麦水溶蛋白的提取工艺优化及其酶解多肽抗氧化活性研究

目的优化复合酶协同超声辅助法提取藜麦水溶蛋白的工艺,探讨7种商业蛋白酶对藜麦多肽的抗氧化活性的影响。方法采用Box-Behnken响应面法,对超声辅助糖化酶-纤维素酶复合酶法提取藜麦水溶蛋白的工艺进行优化。使用碱性蛋白酶、菠萝蛋白酶、风味蛋白酶、木瓜蛋白酶、酸性蛋白酶、胰蛋白酶和胃蛋白酶制备藜麦多肽,通过1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)法和2,2’-联氮二(3-乙基-苯并噻唑-6-磺酸)二铵盐[2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)ammonium salt,ABTS]法对其抗氧化活性进行探究。结果通过响应面模型对复合酶超声辅助提取条件进行优化,得到最佳提取条件为:总加酶量400 U/g、pH 5.0、时间60 min。在此条件下,藜麦水溶蛋白的提取率为75.09 mg/g。藜麦水溶蛋白在木瓜蛋白酶酶解4 h获得的藜麦多肽抗氧化活性最佳,DPPH自由基和ABTS阳离子自由基清除率分别为79.64%和76.14%。结论采用响应面法优化复合酶辅助超声法提取藜麦蛋白的工艺优化方法可行;藜麦蛋白水解物具有较好的抗氧化活性,为藜麦作为功能食品深入开发提供依据。

藜麦FLS基因家族的鉴定、表达及DNA变异分析

【目的】黄酮醇合成酶(FLS)是石竹目植物中多酚类次生代谢的关键酶,为探究FLS基因在藜麦生长发育中的功能,对FLS基因家族进行鉴定和表达分析。【方法】利用生物信息学分析网站,鉴定出CqFLS家族成员,分析其基因结构、蛋白的理化性质、二级结构、三级结构、启动子顺式元件以及系统进化关系等,并通过基因克隆、构建表达载体的方法分析其蛋白表达情况。【结果】共鉴定出3个CqFLSs基因,不均匀分布在2条染色体上,CqFLSs启动子区域包含水杨酸、脱落酸、茉莉酸甲酯和干旱诱导等元件;CqFLS2.1g发现多处InDel和SNP变异,在ch0128871893、28871125、28872881处编码核苷酸删除,且均注释为上游效应,未检测到移码突变;FLS家族系统进化树分析可知,与其他两个基因相比,CqFLS2.1g与CqFLS1.1g、CqFLS3.10g处于不同分支,表达水平也存在差异,CqFLS2.1g可能发生分化;同时,基因表达分析表明,3个CqFLSs基因在青白1号籽粒中整体表达量高于青黑1号和贡扎4号。对克隆出的CqFLS1.1g用0.3 mmol/L的IPTG诱导,在20℃和37℃的条件下均可成功表达。【结论】CqFLS1.1g在花的形成以及籽粒发育过程中发挥作用,而CqFLS2.1g则主要参与藜麦籽粒的形成,CqFLS的表达具有组织特异性,在藜麦生长发育过程中发挥重要的作用。

霜霉病菌胁迫下藜麦内参基因的筛选及其稳定性验证

【目的】筛选藜麦在霜霉病菌胁迫下稳定表达的内参基因,为深入开展藜麦抗霜霉病菌相关基因表达的研究提供依据。【方法】通过实时荧光定量PCR技术以及geNorm、NormFinder、BestKeeper和RefFinder软件分析和评价8个候选内参基因在藜麦霜霉病菌胁迫下的表达稳定性,并通过对CqSGAT基因表达进行分析,进一步验证候选内参基因的稳定性。【结果】geNorm、NormFinder、BestKeeper和RefFinder软件分析结果均显示,藜麦在霜霉病菌胁迫下稳定性较好的内参基因为CqEF-1a、CqMON1和CqRPS18;geNorm软件分析结果显示,藜麦在霜霉病菌胁迫下最适候选内参基因为CqEF-1a和CqRPS18;以CqSGAT为目标基因对候选内参基因进行验证,发现CqMON1不适合作为藜麦在霜霉病菌胁迫下的内参基因。【结论】藜麦在霜霉病菌胁迫下的最适内参基因为CqEF-1a和CqRPS18。