Seed Structure and Localization of Reserves inChenopodium quinoa

The three areas of food reserves in quinoa seeds are: a largecentral perisperm, a peripheral embryo and a one to two-celllayered endosperm surrounding the hypocotyl-radicle axis ofthe embryo. Cytochemical and ultrastructural analysis revealedthat starch grains occupy the cells of the perisperm, whilelipid bodies, protein bodies with globoid crystals of phytin,and proplastids with deposits of phytoferritin are the storagecomponents of the cells of the endosperm and embryo tissues.EDX analysis of the endosperm and embryo protein bodies revealedthat globoid crystals contain phosphorus, potassium and magnesium.These results are compared with studies on other perispermousseeds published to date.Copyright 1998 Annals of Botany Company Chenopodium quinoa,EDX analysis, phytoferritin, phytin, protein bodies, quinoa, seed structure, seed reserves, starch grains.     

Effect of salinity on composition, viability and germination of seeds of Chenopodium quinoa Willd

Salinity influences plant growth, seed yield and seed quality even of halophytic crops such as Chenopodium quinoa. Plant growth, total seed yield, number of seeds, fresh weight and dry weight of seeds, were all significantly reduced in the presence of salinity. Only at high salinity did the content of proteins (as well as total N) increase significantly in the seeds whereas the content of total carbohydrates (as well as total C) decrease. Aside from that the capacity for germination was diminished by a reduced seed size and a disproportionate reduction of the volume of the perisperm. However, the reduced capacity seemed to be compensated by an accelerated germination owing to high Na and Cl concentrations leading to a low water potential in the walls of the plant ovary. At high salinity the passage of NaCl to the seed interior was hindered by the seed cover. There was an obvious gradient between potentially toxic (Na and Cl) and essentially needed elements (K, Mg, Ca, P and S) across the seed coat of salt treated plants and also a significant change of the distribution of elements in the embryo. The results indicate a highly protected seed interior leading to a high salinity resistance of quinoa seeds.     

Assessment of genetic diversity patterns in Chilean quinoa (Chenopodium quinoa Willd.) germplasm using multiplex fluorescent microsatellite markers

Quinoa (Chenopodium quinoa Willd.) is a staple seed crop in the Andean region of South America. Improving quinoa productivity is a primary food-security issue for this region, and has been part of the impetus for the establishment of several new quinoa breeding programs throughout the Andean region. Chilean quinoa has been characterized as morphologically diverse and bifurcated into coastal and highland ecotypes. The success of emerging breeding programs will rely heavily on the development of core germplasm collections and germplasm evaluation—especially of the coastal quinoa ecotypes that are often neglected in traditional breeding programs. Thus, the objective of this study was to characterize and quantify the genetic diversity within 28 Altiplano and 31 coastal Chilean accessions of quinoa using microsatellite markers. To facilitate the analysis, we also report the development of seven sets of fluorescent multiplexed microsatellite PCR reactions that result in genetic information for 20 highly polymorphic microsatellite loci. A total of 150 alleles were detected among the quinoa accession, ranging from 2 to 20 alleles per locus and an average 7.5 allele/locus. Both cluster (UPGMA) and principal component analyses separated the accessions into two discrete groups. The first group contained quinoa accessions from the north (Andean highlands) and the second group consisted of accessions from the south (lowland or coastal). Three accessions from Europe were classified into the southern quinoa group. The data obtained in the diversity analyses highlights the relationships within and among northern and southern Chilean quinoa accessions and provides the quinoa scientific community with a new set of easy to use and highly informative genetic markers.     

Detection and subcellular localization of dehydrin-like proteins in quinoa (Chenopodium quinoa Willd.) embryos

The aim of this study was to characterize the dehydrin content in mature embryos of two quinoa cultivars, Sajama and Baer La Unión. Cultivar Sajama grows at 3600-4000 m altitude and is adapted to the very arid conditions characteristic of the salty soils of the Bolivian Altiplano, with less than 250 mm of annual rain and a minimum temperature of -1 degrees C. Cultivar Baer La Unión grows at sea-level regions of central Chile and is adapted to more humid conditions (800 to 1500 mm of annual rain), fertile soils, and temperatures above 5 degrees C. Western blot analysis of embryo tissues from plants growing under controlled greenhouse conditions clearly revealed the presence of several dehydrin bands (at molecular masses of approximately 30, 32, 50, and 55 kDa), which were common to both cultivars, although the amount of the 30 and 32 kDa bands differed. Nevertheless, when grains originated from their respective natural environments, three extra bands (at molecular masses of approximately 34, 38, and 40 kDa), which were hardly visible in Sajama, and another weak band (at a molecular mass of approximately 28 kDa) were evident in Baer La Unión. In situ immunolocalization microscopy detected dehydrin-like proteins in all axis and cotyledon tissues. At the subcellular level, dehydrins were detected in the plasma membrane, cytoplasm and nucleus. In the cytoplasm, dehydrins were found associated with mitochondria, rough endoplasmic reticulum cisternae, and proplastid membranes. The presence of dehydrins was also recognized in the matrix of protein bodies. In the nucleus, dehydrins were associated with the euchromatin. Upon examining dehydrin composition and subcellular localization in two quinoa cultivars belonging to highly contrasting environments, we conclude that most dehydrins detected here were constitutive components of the quinoa seed developmental program, but some of them (specially the 34, 38, and 40 kDa bands) may reflect quantitative molecular differences associated with the adaptation of both cultivars to contrasting environmental conditions.     

Changes in soluble carbohydrates and related enzymes induced by low temperature during early developmental stages of quinoa (Chenopodium quinoa) seedlings

Low temperature represents one of the principal limitations in species distribution and crop productivity. Responses to chilling include the accumulation of simple carbohydrates and changes in enzymes involved in their metabolism. Soluble carbohydrate levels and invertase, sucrose synthase (SS), sucrose-6-phosphate synthase (SPS) and alpha-amylase activities were analysed in cotyledons and embryonic axes of quinoa seedlings grown at 5 degrees C and 25 degrees C in the dark. Significant differences in enzyme activities and carbohydrate levels were observed. Sucrose content in cotyledons was found to be similar in both treatments, while in embryonic axes there were differences. Invertase activity was the most sensitive to temperature in both organs; however, SS and SPS activities appear to be less stress-sensitive. Results suggest that 1) metabolism in germinating perispermic seeds would be different from endospermic seeds, 2) sucrose futile cycles would be operating in cotyledons, but not in embryonic axes of quinoa seedlings under our experimental conditions, 3) low temperature might induce different regulatory mechanisms on invertase, SS and SPS enzymes in both cotyledons and embryonic axes of quinoa seedlings, and 4) low temperature rather than water uptake would be mainly responsible for the changes observed in carbohydrate and related enzyme activities.     

Construction of a quinoa (Chenopodium quinoa Willd.) BAC library and its use in identifying genes encoding seed storage proteins

Quinoa (Chenopodium quinoa Willd.) is adapted to the harsh environments of the Andean Altiplano region. Its seeds have a well-balanced amino acid composition and exceptionally high protein content with respect to human nutrition. Quinoa grain is a staple in the diet of some of the most impoverished people in the world. The plant is an allotetraploid displaying disomic inheritance (2n=4x=36) with a di-haploid genome of 967 Mbp (megabase pair), or 2C=2.01 pg. We constructed two quinoa BAC libraries using BamHI (26,880 clones) and EcoRI (48,000 clones) restriction endonucleases. Cloned inserts in the BamHI library average 113 kb (kilobase) with approximately 2% of the clones lacking inserts, whereas cloned inserts in the EcoRI library average 130 kb and approximately 1% lack inserts. Three plastid genes used as probes of high-density arrayed blots of 73,728 BACs identified approximately 2.8% of the clones as containing plastid DNA inserts. We estimate that the combined quinoa libraries represent at least 9.0 di-haploid nuclear genome equivalents. An average of 12.2 positive clones per probe were identified with 13 quinoa single-copy ESTs as probes of the high-density arrayed blots, suggesting that the estimate of 9.0× coverage of the genome is conservative. Utility of the BAC libraries for gene identification was demonstrated by probing the library with a partial sequence of the 11S globulin seed storage protein gene and identifying multiple positive clones. The presence of the 11S globulin gene in four of the clones was verified by direct comparison with quinoa genomic DNA on a Southern blot. Besides serving as a useful tool for gene identification, the quinoa BAC libraries will be an important resource for physical mapping of the quinoa genome.     

西藏昆诺阿藜种子可培养内生真菌多样性

探究西藏不同地区昆诺阿藜种子内生真菌的物种组成、多样性等群落结构特征,丰富昆诺阿藜种子相关微生物的可利用资源,对于微生物资源利用和昆诺阿藜病害的生物防控具有重要意义。本研究收集西藏不同海拔3个地区(日喀则、拉萨和林芝)的昆诺阿藜种子,利用传统分离方式进行了真菌的培养。从46份昆诺阿藜种子中共分离出947株真菌,经ITS序列分析结合形态观察鉴定为1门9目12科26属77种。所有的真菌都归类为子囊菌门Ascomycota,菌株数量相对丰度最高的3个属为链格孢属Alternaria (40.2%)、镰刀菌属Fusarium (17.4%)和茎点霉属Phoma (13.9%)。按照多样性指数,日喀则地区的昆诺阿藜内生可培养真菌的多样性明显高于拉萨和林芝两个地区。     

Low-temperature scanning electron microscopy of hyphal colonisation of Chenopodium quinoa during fermentation with Rhizopus oligosporus

In a study of tempe made by fermenting quinoa (Chenopodium quinoa Willd.) with Rhizopus oligosporus Saito, the interaction between the substrate and the fungus varied according to the raw material used. Low-temperature scanning electron microscopy confirmed that hyphal infiltration in quinoa was normally limited by the seed coat, but a massive penetration was found in fragments of quinoa where the testa was broken. This finding has implications for the texture and quality of tempe fermentation of grains and pulses.     

Salt tolerance mechanisms in quinoa (Chenopodium quinoa Willd.)

In the face of diminishing fresh water resources and increasing soil salinisation it is relevant to evaluate the potential of halophytic plant species to be cultivated in arid and semi-arid regions, where the productivity of most crop plants is markedly affected. Quinoa is a facultative halophytic plant species with the most tolerant varieties being able to cope with salinity levels as high as those present in sea water. This characteristic has aroused the interest in the species, and a number of studies have been performed with the aim of elucidating the mechanisms used by quinoa in order to cope with high salt levels in the soil at various stages of plant development. In quinoa key traits seem to be an efficient control of Na⁺ sequestration in leaf vacuoles, xylem Na⁺ loading, higher ROS tolerance, better K⁺ retention, and an efficient control over stomatal development and aperture. The purpose of this review is to give an overview on the existing knowledge of the salt tolerance of quinoa, to discuss the potential of quinoa for cultivation in salt-affected regions and as a basis for further research in the field of plant salt tolerance.     

Lysine biosynthesis and nitrogen metabolism in quinoa (Chenopodium quinoa): study of enzymes and nitrogen-containing compounds.

Aspartate kinase (AK, EC 2.7.2.4), homoserine dehydrogenase (HSDH, EC 1.1.1.3) and dihydrodipicolinate synthase (DHDPS, EC 4.2.1.52) were isolated and partially purified from immature Chenopodium quinoa Willd seeds. Enzyme activities were studied in the presence of the aspartate-derived amino acids lysine, threonine and methionine and also the lysine analogue S-2-aminoethyl-l-cysteine (AEC), at 1 mM and 5 mM. The results confirmed the existence of, at least, two AK isoenzymes, one inhibited by lysine and the other inhibited by threonine, the latter being predominant in quinoa seeds. HSDH activity was also shown to be partially inhibited by threonine, whereas some of the activity was resistant to the inhibitory effect, indicating the presence of two isoenzymes, one resistant and another sensitive to threonine inhibition. Only one DHDPS isoenzyme highly sensitive to lysine inhibition was detected. The results suggest that the high concentration of lysine observed in quinoa seeds is possibly due to a combined effect of increased lysine synthesis and accumulation in the soluble form and/or as protein lysine. Nitrogen assimilation was also investigated and based on nitrate content, nitrate reductase activity, amino acid distribution and ureide content, the leaves were identified as the predominant site of nitrate reduction in this plant species. The amino acid profile analysis in leaves and roots also indicated an important role of soluble glutamine as a nitrogen transporting compound.     

Simple sequence repeat marker development and genetic mapping in quinoa (Chenopodium quinoa Willd.)

Quinoa is a regionally important grain crop in the Andean region of South America. Recently quinoa has gained international attention for its high nutritional value and tolerances of extreme abiotic stresses. DNA markers and linkage maps are important tools for germplasm conservation and crop improvement programmes. Here we report the development of 216 new polymorphic SSR (simple sequence repeats) markers from libraries enriched for GA, CAA and AAT repeats, as well as 6 SSR markers developed from bacterial artificial chromosome-end sequences (BES-SSRs). Heterozygosity (H) values of the SSR markers ranges from 0.12 to 0.90, with an average value of 0.57. A linkage map was constructed for a newly developed recombinant inbred lines (RIL) population using these SSR markers. Additional markers, including amplified fragment length polymorphisms (AFLPs), two 11S seed storage protein loci, and the nucleolar organizing region (NOR), were also placed on the linkage map. The linkage map presented here is the first SSR-based map in quinoa and contains 275 markers, including 200 SSR. The map consists of 38 linkage groups (LGs) covering 913 cM. Segregation distortion was observed in the mapping population for several marker loci, indicating possible chromosomal regions associated with selection or gametophytic lethality. As this map is based primarily on simple and easily-transferable SSR markers, it will be particularly valuable for research in laboratories in Andean regions of South America.     

Soil drenching of paclobutrazol: An efficient way to improve quinoa performance under salinity

Salinity extent and severity is rising because of poor management practices on agricultural lands, possibility lies to grow salt‐tolerant crops with better management techniques. Therefore, a highly nutritive salt‐tolerant crop quinoa with immense potential to contribute for future food security was selected for this investigation. Soil drenching of paclobutrazol (PBZ; 20 mg l^?1) was used to understand the ionic relations, gaseous exchange characteristics, oxidative defense system and yield under saline conditions (400 mM NaCl) including normal (0 mM NaCl) and no PBZ (0 mg l^?1) as controls. The results revealed that salinity stress reduced the growth and yield of quinoa through perturbing ionic homeostasis with the consequences of overproduction of reactive oxygen species (ROS), oxidative damages and reduced photosynthesis. PBZ improved the quinoa performance through regulation of ionic homeostasis by decreasing Na⁺, Cl^?, while improving K⁺, Mg²⁺ and Ca²⁺ concentration. It also enhanced the antioxidative system including ascorbic acid, phenylalanine ammonia‐lyase, polyphenol oxidase and glutathione peroxidase, which scavenged the ROS (H₂O₂ and O₂^?‐) and lowered the oxidative damages (malondialdehyde level) under salinity in roots and more specifically in leaf tissues. The photosynthetic rate and stomatal conductance consequently improved (16 and 21%, respectively) in salt‐stressed quinoa PBZ‐treated compared to the non‐treated ones and contributed to the improvement of panicle length (33%), 100‐grain weight (8%) and grain yield (38%). Therefore, PBZ can be opted as a shotgun approach to improve quinoa performance and other crops under high saline conditions.     

Fermentation of quinoa and wheat slurries by Lactobacillus plantarum CRL 778: proteolytic activity

Quinoa fermentation by lactic acid bacteria (LAB) is an interesting alternative to produce new bakery products with high nutritional value; furthermore, they are suitable for celiac patients because this pseudo-cereal contains no gluten. Growth and lactic acid production during slurry fermentations by Lactobacillus plantarum CRL 778 were greater in quinoa (9.8 log?cfu/mL, 23.1 g/L) than in wheat (8.9 log?cfu/mL, 13.9 g/L). Lactic fermentation indirectly stimulated flour protein hydrolysis by endogenous proteases of both slurries. However, quinoa protein hydrolysis was faster, reaching 40–100 % at 8 h of incubation, while wheat protein hydrolysis was only 0–20 %. In addition, higher amounts of peptides (24) and free amino acids (5 g/L) were determined in quinoa compared to wheat. Consequently, greater concentrations (approx. 2.6-fold) of the antifungal compounds (phenyllactic and hydroxyphenyllactic acids) were synthesized from Phe and Tyr in quinoa by L. plantarum CRL 778, an antifungal strain. These promising results suggest that this LAB strain could be used in the formulation of quinoa sourdough to obtain baked goods with improved nutritional quality and shelf life, suitable for celiac patients.     

Triterpene saponins from Chenopodium quinoa Willd

Twenty triterpene saponins (1-20) have been isolated from different parts of Chenopodium quinoa (flowers, fruits, seed coats, and seeds) and their structures have been elucidated by analysis of chemical and spectroscopic data including 1D- and 2D-NMR. Four compounds (1-4) were identified: 3beta-[(O-beta-d-glucopyranosyl-(1-->3)-alpha-l-arabinopyranosyl)oxy]-23-oxo-olean-12-en-28-oic acid beta-d-glucopyranoside (1), 3beta-[(O-beta-d-glucopyranosyl-(1-->3)-alpha-l-arabinopyranosyl)oxy]-27-oxo-olean-12-en-28-oic acid beta-d-glucopyranoside (2), 3-O-alpha-l-arabinopyranosyl serjanic acid 28-O-beta-d-glucopyranosyl ester (3), and 3-O-beta-d-glucuronopyranosyl serjanic acid 28-O-beta-d-glucopyranosyl ester (4). The following known compounds have not previously been reported as saponin constituents from the flowers and the fruits of this plant: two bidesmosides of serjanic acid (5,6), four bidesmosides of oleanolic acid (7-10), five bidesmosides of phytolaccagenic acid (11-15), four bidesmosides of hederagenin (16-19), and one bidesmoside of 3beta,23,30-trihydroxy olean-12-en-28-oic acid (20). The cytotoxicity of these saponins and their aglycones was tested in HeLa cells. Induction of apoptosis in Caco-2 cells by bidesmosidic saponins 1-4 and their aglycones I-III was determined by flow cytometric DNA analysis. The saponins with an aldehyde group were most active. The relationships between structure and cytotoxic activity of saponins and their aglycones are discussed.     

Ricinosomes provide an early indicator of suspensor and endosperm cells destined to die during late seed development in quinoa (Chenopodium quinoa)

Background and Aims

In mature quinoa (Chenopodium quinoa) seeds, the lasting endosperm forms a micropylar cone covering the radicle. The suspensor cells lie within the centre of the cone. During the final stage of seed development, the cells of the lasting endosperm accumulate protein and lipids while the rest are crushed and disintegrated. Both the suspensor and endosperm die progressively from the innermost layers surrounding the embryo and extending towards the nucellar tissue. Ricinosomes are endoplasmic reticulum-derived organelles that accumulate both the pro-form and the mature form of cysteine endopeptidase (Cys-EP), first identified in castor bean (Ricinus communis) endosperm during germination. This study sought to identify associations between the presence of ricinosomes and programmed cell death (PCD) hallmarks in suspensor and endosperm cells predestined to die during quinoa seed development.

Methods

A structural study using light microscopy and transmission electron microscopy was performed. To detect the presence of Cys-EP, both western blot and in situ immunolocalization assays were carried out using anti-R. communis Cys-EP antibody. A TUNEL assay was used to determine DNA fragmentation.

Results and Conclusions

Except for the one or two cell layers that constitute the lasting endosperm in the mature seed, ricinosomes were found in suspensor and endosperm cells. These cells were also the site of morphological abnormalities, including misshapen and fragmented nuclei, vesiculation of the cytosol, vacuole collapse and cell wall disorganization. It is proposed that, in suspensor and endosperm cells, the early detection of Cys-EP in ricinosomes predicts the occurrence of PCD during late seed development.     

施氮深度和水分胁迫对藜麦幼苗生理及产量的影响

为了探讨藜麦应对施肥深度和水分胁迫的响应,该文以藜麦(Chenopodium quinoa)为材料,在盆栽条件下,设置3种施氮处理[D1(控释尿素施在0～8 cm深度)、D2(控释尿素施在8～16 cm深度)、D3(控释尿素施在16～24 cm深度)]和3种水分处理[W1(正常供水)、W2(中度干旱)、W3(重度干旱)],分析施氮深度和水分胁迫对藜麦幼苗生理及产量的影响。结果表明：(1)相同水分条件下,随着施肥深度的增加,藜麦生长指标(株高、茎粗、叶面积、地上部生物量、主根长、根系表面积、根系体积)、生理指标[超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性、过氧化氢酶(CAT)活性、可溶性糖含量、可溶性蛋白含量、叶绿素总量]和产量指标呈先升高后降低趋势。D2处理(适当的深施氮肥)均高于D1处理(浅层施氮)和D3处理(底层施氮)。(2)相同施氮深度条件下,随着干旱胁迫程度的增加,藜麦生长指标和产量指标呈逐渐降低的趋势,生理指标均呈先升高后降低的趋势。说明藜麦幼苗对水分需求明显,可通过增加抗氧化酶活性和渗透调节物质适应一定程度的干旱,生产实践中应注意苗期水分的供应,以促进生育后期产...     

Phenological growth stages of quinoa (Chenopodium quinoa) based on the BBCH scale

Quinoa (Chenopodium quinoa) is a pseudocereal native from the Andean region of South America that has increased in importance worldwide. Quinoa is now considered an alternative to traditional crops in a climate change scenario, considering its ability to adapt to marginal soils, droughts and frosts. Despite the interesting agronomic and nutritional features of this crop, research into quinoa is characterised by individual attempts to define its phenological stages without an international consensus. A unique criterion to quantify the phenology of quinoa could become a useful tool for researchers and plant breeders in future work by standardising this information for international cooperation. In this article, a proposed scale of the phenological growth stages of quinoa based on the BBCH coding system (Biologische Bundesanstalt Bundessortenamt und CHemische Industrie) was developed. Growth stages were described utilising the decimal code of the BBCH system, and figures were included for the most representative stages.     

Phytochemical and genetic characterization of five quinoa (<Emphasis Type="Italic">Chenopodium quinoa</Emphasis> Willd.) genotypes introduced to Egypt

Due to its substantial nutritional value, quinoa (Chenopodium quinoa Willd.) is currently attracting worldwide attention. Quinoa is characterized by a high adaptability to various environmental conditions. This is the first report on the phytochemical and genetic evaluation of quinoa germplasms introduced to Egypt, and the results could be used to implement propagation techniques in the future. For phytochemical characterization, 41 traits, including primary and secondary metabolites, antioxidant molecules, sugars, organic acids and fatty acids, were evaluated. At the same time, 4 RAPD and 7 ISSR markers were used for genetic analysis. UPGMA analysis of RAPD and ISSR polymorphic markers, their combined dataset and phytochemical traits were used to evaluate genetic relationships among genotypes. The quinoa genotypes displayed reasonable variation in the studied phytochemical traits. The results of the genetic analysis confirmed that RAPD and ISSR markers could be used to distinguish effectively quinoa genotypes. The phytochemical and genetic characterization reported herein will be a promising guide for breeding seed quality in quinoa.     

A dopamine-based biosynthetic pathway produces decarboxylated betalains in Chenopodium quinoa

Betalains are the nitrogenous pigments that replace anthocyanins in the plant order Caryophyllales. Here, we describe unconventional decarboxylated betalains in quinoa (Chenopodium quinoa) grains. Decarboxylated betalains are derived from a previously unconsidered activity of the 4,5-DOPA-extradiol-dioxygenase enzyme (DODA), which has been identified as the key enzymatic step in the established biosynthetic pathway of betalains. Here, dopamine is fully characterized as an alternative substrate of the DODA enzyme able to yield an intermediate and structural unit of plant pigments: 6-decarboxy-betalamic acid, which is proposed and described. To characterize this activity, quinoa grains of different colors were analyzed in depth by chromatography, time-of-flight mass spectrometry, and reactions were performed in enzymatic assays and bioreactors. The enzymatic-chemical scheme proposed leads to an uncharacterized family of 6-decarboxylated betalains produced by a hitherto unknown enzymatic activity. All intermediate compounds as well as the final products of the dopamine-based biosynthetic pathway of pigments have been unambiguously determined and the reactions have been characterized from the enzymatic and functional perspectives. Results evidence a palette of molecules in quinoa grains of physiological relevance and which explain minor betalains described in plants of the Caryophyllales order. An entire family of betalains is anticipated.

A biosynthetic pathway produces unconventional plant pigments betalains derived from dopamine in quinoa.