Characterization of Agro-diversity by Seed Storage Protein Electrophoresis:Focus on Rice Germplasm from Uttarakhand Himalaya,India

The characteristics of 48 rice varieties from Uttarakhand Himalaya, India were detected by morphological and biochemical markers. The grains of the selected rice varieties varied in their morphological （grain length, grain width and grain weight） and biochemical characters （sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE）. Based on the presence of 70, 65, 60, 57, 37-39, 22-23, 13 and 10 kDa protein bands in the 48 rice varieties, seven types of profiles were identified. An unweighted pair group average method with arithmetic mean （UPGMA） dendrogram based on cluster analysis of genetic similarity of the protein bands showed two distinct groups with 1%-78% similarity coefficients. The presence of characteristic bands in selected varieties is a useful parameter for identification of rice germplasm.

A Dominant Locus, qBSC-1, Controls β Subunit Content of Seed Storage Protein in Soybean(Glycine max(L.) Merri.)

Soybean seed storage protein is one of the most important plant vegetable proteins, and β subunit is of great significance to enhance soybean protein quality and processing property. F2 segregated population and residual heterozygous lines（RHL） derived from the cross between Yangyandou（low level of β subunit） and Zhonghuang 13（normal level of β subunit） were used for mapping of β subunit content. Our results showed that β subunit content was controlled by a single dominant locus, qBSC-1（β subunit content）, which was mapped to a region of 11.9 cM on chromosome 20 in F2 population of 85 individuals. This region was narrowed down to 2.5 cM between BARCSOYSSR_20_0997 and BARCSOYSSR_20_0910 in RHL with a larger population size of 246 individuals. There were 48 predicted genes within qBSC-1 region based on the reference genome（Glyma 1.0, Williams 82）, including the two copies of β subunit coding gene CG4. An InDel marker developed from a thymine（TT） insertion in one copy of CG4 promoter region in Yangyandou cosegregrated with BARCSOYSSR_20_0975 within qBSC-1 region, suggesting that this InDel marker maybe useful for marker-assisted selection（MAS）.

New combination in Veronica (Scrophulariaceae s.l.) based on morphological characters and the seed storage protein polymorphism

Veronica erciyasdagi （M. A. Fischer） C. Vural comb. ＆ stat. nov., previously regarded as a variety, is proposed as a new combination, based on the morphological characters and seed storage protein polymorphism presented in this study. In addition, information about the ecology and conservation status of Veronica erciyasdagi was reported. This taxon is endemic to central Anatolia, Turkey and is critically endangered.

Orchestrating seed storage protein and starch accumulation toward overcoming yield–quality trade-off in cereal crops

Achieving high yield and good quality in crops is essential for human food security and health.However,there is usually disharmony between yield and quality.Seed storage protein(SSP)and starch,the predominant components in cereal grains,determine yield and quality,and their coupled synthesis causes a yield–quality trade-off.Therefore,dissection of the underlying regulatory mechanism facilitates simultaneous improvement of yield and quality.Here,we summarize current findings about the synergistic molecular machinery underpinning SSP and starch synthesis in the leading staple cereal crops,including maize,rice and wheat.We further evaluate the functional conservation and differentiation of key regulators and specify feasible research approaches to identify additional regulators and expand insights.We also present major strategies to leverage resultant information for simultaneous improvement of yield and quality by molecular breeding.Finally,future perspectives on major challenges are proposed.

Regulation of seed storage protein synthesis in monocot and dicot plants:A comparative review

Seeds are a major source of nutrients for humans and animal livestock worldwide.With improved living standards,high nutritional quality has become one of the main targets for breeding.Storage protein content in seeds,which is highly variable depending on plant species,serves as a pivotal criterion of seed nutritional quality.In the last few decades,our understanding of the molecular genetics and regulatory mechanisms of storage protein synthesis has greatly advanced.Here,we systematically and comprehensively summarize breakthroughs on the conservation and divergence of storage protein synthesis in dicot and monocot plants.With regard to storage protein accumulation,we discuss evolutionary origins,developmental processes,characteristics of main storage protein fractions,regulatory networks,and genetic modifications.In addition,we discuss potential breeding strategies to improve storage protein accumulation and provide perspectives on some key unanswered problems that need to be addressed.

Longevity of Juniperus procera seed lots under different storage conditions: implications for ex situ conservation in seed banks

Juniperus procera Endl. is economically important timber species, but its populations are extremely small and fragmented in its natural habitat, thus, calling for immediate ex situ conservation. Here we examined the effects of seed sources and storage temperature on the longevity of Juniperus procera seed lots through collection and preservation of seeds in seed banks. Seeds were collected from nine sites across the species natural distribution in Ethiopia and stored in four warehouses： modern cold room （5℃）, mud house （15℃）, concrete block house （17℃ or corrugated iron house （20℃） for 42 months. Every three months, a random sample of stored seeds were drawn and tested for germination. A highly significant variation （p 0.01） in germination of stored seeds was observed among different storage environments, seed lots, and duration of storage. Over the storage period, seeds stored in the cold room had the highest mean percentage germination, followed by the mud house, corrugated house and blocket house. The cold room （41%） and the mud house （38%） maintained the same level of germination as the intitial germination of the seedlotds （42%）. The variation in longevity of stored seeds was significnatly correlated with the initial germination of seed lots （r 0.80; p 0.01）. Cold storage also resulted in enhancement of germination through its stratification effect that terminated the non-deepphysiological dormancy of juniper seeds. In conclusion, seed lots with good initial germination can be effectively stored in cold room （5℃） up to four years. In the absence of modern cold stores, mud houses can be used as a good alternative to store seeds at local level.

Rice seed storage proteins:Biosynthetic pathways and the effects of environmental factors

Rice(Oryza sativa L.)is the most important food crop for at least half of the world’s population.Due to improved living standards,the cultivation of high-quality rice for different purposes and markets has become a major goal.Rice quality is determined by the presence of many nutritional components,including seed storage proteins(SSPs),which are the second most abundant nutrient components of rice grains after starch.Rice SSP biosynthesis requires the participation of multiple organelles and is influenced by the external environment,making it challenging to understand the molecular details of SSP biosynthesis and improve rice protein quality.In this review,we highlight the current knowledge of rice SSP biosynthesis,including a detailed description of the key molecules involved in rice SSP biosynthetic processes and the major environmental factors affecting SSP biosynthesis.The effects of these factors on SSP accumulation and their contribution to rice quality are also discussed based on recent findings.This recent knowledge suggests not only new research directions for exploring rice SSP biosynthesis but also innovative strategies for breeding high-quality rice varieties.

Seed Germinability of 23 Crop Species After a Decade of Storage in the National Genebank of China

Seed viability of 18 576 accessions of 23 crops was monitored. After 10-12 years storage in National Crop Genebank ofChina (NCGC), more than 96.12% of the accessions maintained high germination percentage (>85%). The germination of95 accessions, accounting for 0.51% of the total, declined significantly from above 80% to below 70%. For each crop ofcarrot (Daucus carota var. sativa DC.), lettuce (Lactuca sativa L.), cotton (Gossypium sp. L.), flax (Linum usitatissimumL.) and castor-oil plant (Ricinus communis L.), the mean monitored germination percentage declined significantly as awhole, of which carrot and lettuce seeds lost viability more rapidly. Seed initial germination percentage and pre-storageenvironments affect subsequent seed viability in storage.

Effect of Moisture Content on Melilotus suaveolens Seed Quality During Ultra-drying Storage

In order to enhance the seed storability and supply high-quality seeds for vegetation restoration in the arid and semi-arid regions in Northwest China,the effects of ultra-drying and accelerated aging on the physiological characterstics of Melilotus suaveolens seeds were studied.Melilotus suaveolens seeds were dried in a desiccator containing silica gel to 80,53,42,33,23 and 16 g·kg-1 of moisture contents（MC）,respectively.The parameters of the seed quality including germination energy（GE）,germination percentage（GP）,relative conductivity（RC）,dehydrogenase activity（DA） and α-amylase activity（AA） were determined after ultra-drying and accelerated aging.The results showed that ultra-dried seeds with 42 g·kg-1 of MC showed the minimum changes of GE and GP before and after seed aging.Moreover,ultra-dry seeds with 42 g·kg-1 of MC showed higher DA and AA,and lower RC than non-ultra-drying seeds.Therefore,ultra-drying to 42 g·kg-1 of MC was helpful for M.suaveolens seed storage.

Research of Seed Testa Structure and Storage Material of Peanut Germplasm with Different Resistance to A. flavus

There was an obvious relationship between seed testa structure, storage material and resistance to A. flavus of peanut. Results showed that seed testa of peanut germplasm with high resistance （HR） to A. flavus infection had thicker wax layer, integrated and tight epidermis layer, regular vascular tissue range. However, the seed testa of peanut germplasm with high sensitivity （HS） to A. flavus had the reverse results, and results of those with medium resistance （MR） to A. flavus lay in between, but changes of testa thickness were not significant among different resistance kinds. Results also showed that some seed storage materials were closely related with resistance potential to A. flavus. It seemed that varieties with higher resistance to A. flavus had higher oleic acid and protein content, lower linoleic acid and fat content. Content of palm acid, total sugar and VE did not show positive relationship with the resistance to A. flavus.

Calophyllum inophyllum: recalcitrant or intermediate seed?

We studied seed storage behaviour of a multipurpose tree Calophyllum inophyllum （Clusiaseae）. Seeds were collected at Roslyn bay （2307′60＂ S, 150°43′60＂ E） Central Queensland. Seed drying and desieeation tolerance tests were carried out according to International Seed Testing Association （ISTA） rules. We found C. inophyllum seeds to be recalcitrant and vulnerable to chilling injury, hence, unsuited for cold storage. C. inophyllum seeds maintained their viability for an appreciable period （〉 8 months） if stored in wanner environments.

The Association between Germination Vigour and Biochemical, Physiological, Subcellular Changes in Bambara Groundnut Seeds after Seed Scarification

Seed storage reserves and organelles after scarification(chemical,mechanical,control)of bambara groundnut with different seed coat colours(cream,light brown and brown)were examined in relation to germination vigour.Seed storage proteins,oil and sugar content,starch granules and protein bodies,germination vigour index(GVI)and mean germination time(MGT)were measured.There were significant(p<0.05)effects of seed scarification on protein body diameter,seed oil content and sugar content.Chemical scarification resulted in seeds having larger starch granules,followed by the control and mechanical scarification that resulted in seeds having smaller starch granules.Mechanical scarification had the highest(p>0.05)protein density,followed by chemical scarification and control seeds that had the lowest protein density.Seeds from the control had the highest oil content,followed by seeds from mechanical scarification and seeds from chemical scarification that had the lowest oil content.Seeds from the control and chemical scarification had larger diameter of protein bodies and sugar content.Seed coat colour significantly(p<0.05)influenced the area and diameter of protein bodies,starch granule area,seed sugar content.Brown seeds produced larger protein bodies and the highest oil content,while light brown seeds had larger area of starch granules and the highest sugar content.Seed storage reserves had an influence on bambara groundnut germination vigour.While favouring dark coloured seeds such as light brown and brown,bambara groundnut seed users should adapt chemical seed scarification to enhance germination vigour of this crop.

Dormancy Breaking and Storage Behavior of Garcinia cowa Roxb. （Guttiferae） Seeds： Implications for Ecological Function and Germplasm Conservation

Abstract: The dormancy breaking and storage behavior of Garcinia cowa Roxb. seeds were investigated. The seeds of G. cowa had 8–11 months dormancy in their natural habitat. Seeds were matured and dispersed at the end of the rainy season (mid-late August to late September) and were scatter-hoarded by rodents as food for winter after the seeds had fallen to the ground. Seedlings often emerged in the forest during the rainy season (May to August) the following year. Intact seeds of G. cowa failed to germinate after being sown at 30 °C for 120 d and the mean germination time (MGT) of seeds cultured in a shade (50% sunlight) nursery was 252 d. The most effective method of breaking dormancy was to remove the seed coat totally, which reduced the MGT to 13 d at 30 °C. Germination was also promoted by partial removal of the seed coat (excising the hilum and exposing the radicle) and chemical scarification (immersion in 1% H2O2 for 1 d). Unscarified seeds take up water rapidly in the first 96 h, but water was absorbed by the outside seed coat, without penetrating through it. The moisture content (MC) of G. cowa seeds was high (50% in fresh weight) at shedding. The seeds could tolerate desiccation to some extent, until the MC reached approximately 40%; below that, the viability decreases rapidly and all seeds died at approximately 17% of MC. Seed viability decreased rapidly when seeds were chilled at 4 °C; germination was 2% after storage for 1 week. Even stored at 10 °C, seeds began to be damaged after 4 weeks. Seed storage for 1 yr revealed that in both dry (relative humidity (35 ± 5)%) and moist (wet sand) storage conditions, seed viability declined, but germination percentages for seeds stored under moist conditions are better than for seed stored under dry conditions. Because of their low tolerance to desiccation, marked chilling sensitivity and relatively short lifespan, G. cowa seeds should be classified into the tropical recalcitrant category. The ecological implications of dormant recalcitrant seeds and cues on storing recalcitrant seeds were discussed.

小麦种子在不同贮存条件下保存10年后生活力的差异

冬小麦农大139含水量为6.7%和11.8%。分别密封保存于室温和0～5℃冰箱。10年后,测定其发芽势,发芽率等指标,结果表明:①在室温条件下,含水量为11.8%的种子全部死亡,含水量为6.7%的种子有近50%的发芽率同保存前种子发芽率相比明显下降。②在0～5℃冰箱保存的种子,两种含水量均达到80%左右的发芽率,同保存前相比没有明显差别。统计分析表明,贮存温度及贮存温度和种子含水量的互作对贮存种子活力的影响达到极显著水平。

Genetic Diversity and Classification of Chinese Elite Foxtail Millet [Setaria italica (L.) P. Beauv.] Revealed by Acid-PAGE Prolamin

Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important cereal crops in China. Evaluation of germplasm and genetic diversity of foxtail millet is still in its infancy, but prolamin could play an important role as a protein marker. To investigate the genetic diversity and population structure of foxtail millet from different ecological zones of China, 90 accessions of foxtail millet were collected from three major ecological areas: North, Northwest, and Northeast China. The prolamin contents were examined by acid polyacrylamide gel electrophoresis (acid-PAGE). Five to twenty-two prolamin bands appeared in tested varieties, of which were polymorphic, so prolamin patterns of foxtail millet varieties can be used in variety identification and evaluation. Structure analysis identified six groups, which matches their pedigree information but not their geographic origins. This indicated a high degree (87.78%) of consistency with a phylogenetic classification based on SSR. The results showed prolamin banding patterns were an effective method for analyzing foxtail millet genetic variability.

Disulphide Linkages Occur in Many Polypeptides of Rice Protein Fractions: a Two-dimensional Gel Electrophoretic Study

Most of the work on rice storage proteins focused on the major fraction - glutelin. In contrast to previous reports of the occurrence of single legumin-like polypeptide pair in glutelin, two-dimensional gel electrophoresis revealed many glutelin bands, with some having and some lacking disulphide-linkages. Five legumin-like polypeptide pairs exhibited wide heterogeneity over a range of molecular weight （Mr） 25 to 60 kDa, each having a large subunit （Mr ranged from 18 to 40.5 kDa） disulphide-bonded to a small subunit （Mr ranged from 16.5 to 18.0 kDa）. A band of 49 kDa was homodimeric with two subunits of 29 kDa each; a polypeptide of 51 kDa which altered position to ones corresponding to 53 kDa and 57 kDa on 2-D gels contained intrapolypeptide linkages. Polypeptides of 65 kDa and 60 kDa occurred as aggregates of 110 kDa. The number of polypeptides in other seed protein fractions albumins, globulins and prolamins, varied from four to ten. One of the albumin bands had intrapolypeptide disulphide linkages （20 kDa） and the globulins contained two such bands （13.5 and 20 kDa）. Thus, the present study provides a description of the polypeptide composition of different rice protein fractions that is finely resolved with respect to the occurrence of disulphide linkages.

Effect of Allelic Variation of β-conglycinin α-subunit Locus cgy-2 on Soybean(Glycine max) Quality Ⅰ. Evaluation of Free Amino Acid Content

The effects of the deficiency of the allergenicα-subunit on soybean amino acid(AA)composition were studied using the cultivar Dongnong 47(DN47)as a genetic background.The near-isogenic line(NIL)NIL-DN47-Δα of DN47,with an introgression of theα-null trait allele from the high protein donor parent RiB,was created by marker assisted background selection and used to investigate the AA content and nutritional quality.The contents of crude protein,the total AAs,the total essential amino acids(EAAs)and sulfur-containing(Met and Cys)AAs increased by 4.11%,4.16%,5.20% and 11.96%,respectively in NIL-DN47-Δα compared with DN47.Analyses of the total EAAs(TEAAs)and the EAA index(EAAI)revealed that both parameters in NIL-DN47-Δα were higher than those in DN47.The null-allele of theα-subunit positively affected the AA scores.The quantitative changes in free AAs(FAAs)in the developing seeds of NIL-DN47-Δα and DN47 were compared as of 15 days after flowering(DAF)until maturity.The results showed that the total FAA content in NIL-DN47-Δα was significantly higher than that in the DN47 throughout the late maturation stage(40-60 DAF)of seeds.The high concentration of the FAAs in cgy-2 mutant seeds was a consequence of the high rates of synthesis and/or accumulation of individual FAAs during seed maturation where 25 DAF was an important turning point in the accumulation of the FAAs.The FAA contents of single soybeanα-null,double(α+α′)and triple(α+α′+group I)-null mutant combination lines were investigated.In all of these combinations,introduction of the cgy-2 gene invariably raised the FAA content of mature seeds above that of the DN47.In summary,the enhanced protein quality in cgy-2 mutants resulted from several factors.(1)There was a general increase in the contents of most AAs and FAAs in NIL-DN47-Δα.(2)The induced synthesis of free Arg contributed effectively to the high FAAs of various storage-protein-deficiency mutants.For example,in the S2(null α,group I),the free Arg content was seven times as much as that of DN47,accounting for more than half of the total FAA content in the seed.(3)The increase of sulfur-containing AAs in theα-null type NIL mainly resulted from elevated Met content.These data suggested that the cgy-2 mutation might improve the protein quality of soybean seeds and that lacked of the allergenicα-subunit resulted in increased the FAA content.

Antimicrobial Activity of a Cys-Rich Peptide Derived from a Centrosema virginianum Vicilin

Many antimicrobial peptides (AMPs) have been identified in plants. These peptides are highly divergent at the primary sequence level and vary in their hierarchical structures. Some common biochemical features include the ability to form disulfide bonds, tandemly repeated amino acid sequences and a net charge at pH 7. Unusual Cysteine containing repeats has been identified in several plant seed storage proteins that may act as AMPs. We identified a Cys repeat within a vicilin (seed storage protein) of a wild legume, Centrosema virginianum. Cleavage of the vicilin protein during germination would generate a vicilin derived Cys peptide (VDCP). We investigated the antimicrobial properties of this VDCP and compared its efficacy as an antimicrobial agent to VDCPs from other species. We developed transgenic tobacco plants that expressed cloned sequences encoding the Cysteine repeat unit from C. virginianum, Theobroma cacao and Gossypium hirsutum. Extracts from fully expanded leaves were tested for antimicrobial activity against a fungal pathogen, Botrytis cinerea. The Cys motif from C. virginianum was also expressed in two E. coli cell lines (reducing or oxidizing cytoplasm) and peptide fusion protein fractions were tested for antimicrobial activity against a battery of fungal strains. The unique Cysteine repeat single unit from C. virginianum exhibited antimicrobial properties greater than or equal to the antimicrobial activity associated with expression of the multiple Cys-repeat VDCPs from G. hirsutum or T. cacao in transgenic tobacco. When expressed in bacteria, a C. virginianum VDCP fusion protein exhibited antifungal activity against 3 of the 4 fungi tested. Although the primary role of seed storage proteins is to provide a pool of amino acids and nitrogen for germinating seeds and developing plantlets, it is likely that seed storage protein proteolytic products also provide beneficial antimicrobial properties during germination and young plantlet development.

Starch Mobilization in Ultradried Seed of Maize （Zea mays L.） During Germination

The effects of ultradry storage on the starch mobilization in maize (Zea mays L.) seed after aging were investigated. The results indicated that there were no significant differences in the content of ATP, starch, and soluble sugar, as well as the activity of amylase, between ultradried seeds and seeds stored at ?20 °C during germination. These results were consistent with the higher level of vigor of the ultradried seed. Sieve tube introduction of a fluorescence dye (carboxyl fluoresceindiacetate) and laser confocal microscopy were used to study the development of plasmodesmata in the ultradried seeds. The results indicated that plasmodesmata developed well in ultradried seeds. Fluorescence analysis also showed that the fluorescence intensity in the radicle of ultradried seeds was stronger than that in seeds with a higher moisture content. This suggests that ultradry treatment has no adverse effects on the seeds. After seed imbibition, cell orgaelles could be resumed. It is concluded that ultradry seed storage is beneficial for maintaining seed vigor and that starchy mobilization proceeds regularly during germination.