Construction and Quality Analysis of Full-length cDNA Library of Phyllostachys heterocycla Germinating Seeds

[Objective] This study aimed to construct the full-length cDNA library for ger- minating seeds of Phyllostachys heterocycla [Method] Germinating seeds of P. hetero- cycla were used as experimental materials to construct the full-length cDNA library by using Oligo-capping method. [Result] The constructed library has a total capacity of 6.5×10^6 recombinant clones, and a low proportion of clones without inserted frag- ments; the size of inserted fragments ranges between 0.3-5.0 kb, with strict classifi- cation and ideal consistency. Furthermore, the proportion of clones harboring long in- serted fragments （1.0-5.0 kb） is as high as 30%, achieving the standard for high- quality full-length cDNA library. [Conclusion] The full-length cDNA library of germinat- ing seeds of P. heterocycla was successfully constructed, which laid important foun- dation for the functional genomics research of bamboo plants.

EMS-Mediated Mutagenesis in Marigold Seeds and Its Effects on Seedling Growth and Physiology

Marigolds(Tagetes spp.)are popular horticultural plants worldwide.The current study aimed to investigate the optimal mutagenic conditions for marigold seeds using EMS(ethyl methanesulfonate)mutagenesis.Different con-centrations and treatment times of EMS were applied to investigate their effects on the marigold seed germination rate,growth traits,antioxidant enzyme activities(i.e.,SOD and POD),and malondialdehyde(MDA)contents.Results indicated that with increasing the EMS treatment duration and concentration,the seed germination rate and growth treatments were reduced,accompanied by elevated MDA content.In addition,SOD and POD activ-ities initially correlated positively with the growth tratis at the lowest concentrations and shortest durations of EMS,but such relationship diminished beyond certain thresholds.The comprehensive analysis identified the opti-mal mutagenic conditions as 1%EMS treatment for 12 h,achieving a semi-lethal dose and enhancing stress-resis-tant components in seedlings.Thesefindings are pivotal for advancing genetic enhancement and germplasm innovation in marigolds.

Effects of Nitric Oxide on the Germination of Wheat Seeds and Its Reactive Oxygen Species Metabolisms Under Osmotic Stress

Effects of sodium nitroprusside (SNP), a nitric oxide (NO) donor, on the germination and metabolism of reactive oxygen species were surveyed in wheat (Triticum aestivum L.) seeds. Germination of wheat seeds and even the elongation of radicle and plumule were dramatically promoted by SNP treatments during the germination under osmotic stress. Meanwhile, activities of amylase and EP were enhanced, thus leading to the degradation of storage reserve in seeds. After osmotic stress was removed, higher viability of wheat seeds was also maintained. In addition, the activities of CAT, APX and the content of proline were increased by SNP treatment simultaneously, but activities of LOX were inhibited, and both of which were beneficial for improving the antioxidant capacity during the germination of wheat seeds under osmotic stress. It was also shown that the increase of the activity of amylase induced by SNP in embryoless half-seeds of wheat in the beginning period of germination (6 h) might be indirectly related to GA(3).

Analysis of Amylase and Superoxide Dismutase Isozymes During the Germination Process of Emmenopterys henryi Oliv Seeds

[ Objective] The study was to understand the changes of amylase（AMY） and superoxide dismutase（SOD） isozymes during the ger- mination process of Emmenopterys henryi Oliv seeds. [ Metbod] By employing polyacrylamide gel electrophoresis method, the expressions of AMY and SOD isozymes during seed germination process were analyzed. ~ Result] The main AMY bands remained strong during the whole peri- od and a new band A2 appeared in the middle and late period of seed germination. Some new SOD bands occurred at the early stage, then be- came weak or disappeared in the middle period, and band S6 became intense in the late peried. [ Conclusion.] The expression of AMY and SOD isozyme gene has temporal difference during germination of E. henryi Oliv seeds.

Physiological characteristics changes of Aesculus chinensis seeds during natural dehydration

A study was conducted to determine the physiological characteristics changes of Aesculus chinensis seeds during natural dehydration in 2003. The results showed that A. chinensis seeds were recalcitrant with being highly desiccation-sensitive. The seed moisture content of fresh fruits was higher than 60%. When the seeds were naturally dried for 30 days, their moisture content declined to 30.2% and their viability was completely lost. The seed germination percentage had a small increase at the beginning of desiccation and then decreased rapidly. The relative electrical conductivity of the A. chinensis seeds increased along with a decrease in seed moisture content. However, there was an abnormal increase in relative electrical conductivity when the seed moisture content was between 53.7% and 50.9%. Superoxide dismutase （SOD） activity decreased rapidly in the period of desiccation except for an abnormality when the seed moisture content was between 53.7% and 50.9%. Malondialdehyde （MDA） content increased slowly at the early stage of desiccation and then rose rapidly after the moisture content was below 50.9%. The soluble sugar content in seeds slowly increased with the increasing period of desiccation. The seed germination percentage was at the high level when seed moisture content was in range of 47%- 60%, which suggests that this was the optimum moisture content for maintaining A. chinensis seed viability.

Effects of Pb Stress on Seed Germination and Seedling Growth of Two Weed Species Seeds

Objective] The aim was to investigate the effects of Pb stress on seed germination and seedling growth of Chenopodium glaucum and Chenopodium serot-inum L. [Method] With C. glaucum and C. serotinum as the study objects, the ef-fects of different concentrations of Pb on their seed germination and seedling growth, as wel as on the leaf SOD, POD and CAT activity were studied primarily. [Result] The germination potential of both the two kinds of weed seeds increased first and then decreased with the increase of Pb concentration. The germination po-tential of C. serotinum and C. glaucum seeds reached the peak at Pb concentration of 25 and 50 mg/L, respectively. With the increase of Pb treatment concentration, the germination rate, germination index and vigor index of C. serotinum and C. glaucum seeds decreased, and their bud length and root length also reduced gradu-al y. ln overal , the leaf SOD, POD and CAT activity of C. glaucum and C. serot-inum_ seedlings increased first and then decreased with the increased treatment concentration of Pb. The leaf SOD and POD activity of C. glaucum and C. serot-inum seedlings reached the maximum at Pb concentration of 200 and 100 mg/L, respectively, and the leaf CAT activity reached the maximum at Pb concentration of 100 mg/L. [Conclusion] The two Chenopodium species had strong tolerance to Pb stress, and they could be used as alternative accumulator plants of soil Pb contami-nation.

Effects of Used Battery on Key Enzyme Activity during the Germination of Wheat Seeds

[Objective] This study aimed to explore the effects of used battery lixivium on wheat germination. [Method] The wheat seeds were treated with used battery lix- ivium at different concentrations to detect the change of activities of amylase, pro- tease, pyruvate dehydrogenase （PDH） and polyphenol oxidase （PPO） during the ger- mination period. [Result] The results showed that the used battery affected enzyme activity. With the increase of concentration of used battery lixivium, trends of the changes of amylase and protease activities were not different. The activities were en- hanced at low concentrations of lixivium, while were inhibited at high concentrations. The tends of changes of pyruvate dehydrogenase （PDH） and polyphenol oxidase （PPO） activities were not consistent with that of either amylase or protease, which showed continuous downward trends with the increasing concentration of used battery lixivium. [Conclusion] This study is of great practical significance for understanding the effects of used battery lixivium on the germination of wheat seeds.

Effects of Phosphine on Germination and Physiological Characteristics of Rice Seeds

[Objective] This study aimed to investigate the effects of phosphine on germination and physiological characteristics of rice seeds. [Method] Simulation envi- ronments were conducted to study the effects of high-level phosphine on germination status and physiological characteristics of rice seeds and explore the early environ- mental and ecological effects of phosphine on rice growth in phosphorus cycle of paddy field. [Result] Experimental results showed that the increase of phosphine con- centration in the environment resulted in the decrease of germination rate and ger- mination potential by 11.11% and 19.71%, respectively. In addition, the activities of catalase （CAT） and peroxidase （POD） were reduced to 94.35% and 92.61%, respec- tively; the content of malondialdehyde （MDA） was maximally increased by 29.11%, indicating that both germination potential and growth condition of rice seeds were in- hibited under conditions of high-level phosphine. [Conclusion] This study provided theoretical basis for investigating the effects of phosphine on germination of rice seeds under natural environment.

Influence of Temperature and Salinity on Germination of Eelgrass (Zostera marina L.) Seeds

Seagrass restoration as part of ocean ecosystem protection has been launched for many years all over the world, but intensive research on this subject in China has just begun in recent years. Seed broadcasting has been widely accepted as the most potentially useful method for seagrass restoration over large areas. We examined the influence of key environmental factors on seed germination to help promote eelgrass bed restoration. Under anoxic conditions, the influence of temperature and salinity on the germination rate of eelgrass (Zostera marina L.) seeds was examined at different combinations of four temperatures (4, 9, 14, and 24℃) and nine salinities (5 to 45, increment of 5). The effect of significant interaction of temperature and salinity on germination rate was observed (ANOVA) (P<0.001). The highest germination rate (83.3 ± 3.5)% was reached in 8 weeks at 14℃ and salinity 5. Higher temperature significantly increased the germination rate at salinity 5 (P<0.001) during the whole observation period except for 24℃, while lower salinity significantly increased the germination rate at 14℃ (P<0.001). Although significant interaction was found between temperature and salinity (P<0.001), the influence of salinity was stronger than that of temperature for the germination of eelgrass seeds. These results provide useful information for the propagation of artificial seedlings for seagrass restoration in China.

Does Cold Plasma Affect Breaking Dormancy and Seed Germination? A Study on Seeds of Lamb's Quarters(Chenopodium album agg.)

Low-pressure discharge is applied for stimulation of germination of two seed lots of Lamb＇s Quarters （Chenopodium album agg.） with different starting germinations （17%, 8%） and in different stages of dormancy. Different exposition durations with cold plasma treatment were applied. The variable of the ratio cumulative germination was calculated. The Richards＇ equation was used for curve-fitting and simulation of the growth curves. Population parameters, namely Vi - viability, Me - time, Qu - dispersion, and Sk - skewness, counted from the curves described the germination rate well. Significant differences among Qu confirmed the erratic dormancy and gradual germination of Lamb＇s Quarters. No difference in the Me parameter was found between two tested seed lots, and no interspecies characteristics were changed using low-pressure discharge. The results suggested that plasma treatment changed seed germination in Lamb＇s Quarters seeds.

GABA Enhances Thermotolerance of Seeds Germination by Attenuating the ROS Damage in Arabidopsis

Seeds germination is strictly controlled by environment factor such as high temperature(HT)through altering the balance between gibberellin acid(GA)and abscisic acid(ABA).Gama-aminobutyric acid(GABA)is a small molecule with four-carbon amino acid,which plays a crucial role during plant physiological process associated with pollination,wounding or abiotic stress,but its role in seeds germination under HT remains elusive.In this study we found that HT induced the overaccumulation of ROS,mainly H_(2)O_(2) and O_(2)^(-),to suppress seeds germination,meanwhile,HT also activated the enzyme activity of GAD for the rapid accumulation of GABA,hinting the regulatory function of GABA in con-trolling seeds germination against HT stress.Applying GABA directly attenuated HT-induced ROS accumulation,upregulated GA biosynthesis and downregulated ABA biosynthesis,ultimately enhanced seeds germination.Consistently,genetic analysis using the gad1/2 mutant defective in GABA biosynthesis,or pop2-5 mutant with high endogenous GABA content supported the potential function of GABA in improving seeds germination tolerance to HT through scavenging ROS overaccumulation.Based on these data,we propose that GABA acts as a novel signal to enhance thermotolerance of seeds germination through alleviating the ROS damage to seeds viability.

S-Nitrosoglutathion Reductase Activity Modulates the Thermotolerance of Seeds Germination by Controlling ABI5 Stability under High Temperature

Seed germination or dormancy status is strictly controlled by endogenous phytohormone and exogenous environment signals.Abscisic acid(ABA)is the important phytohormone to suppress seed germination.Ambient high temperature(HT)also suppressed seed germination,or called as secondary seed dormancy,through upregulating ABI5,the essential component of ABA signal pathway.Previous result shows that appropriate nitric oxide(NO)breaks seed dormancy through triggering S-nitrosoglutathion reductase(GSNOR1)-dependent S-nitrosylation modification of ABI5 protein,subsequently inducing the degradation of ABI5.Here we found that HT induced the degradation of GSNOR1 protein and reduced its activity,thus accumulated more reactive nitrogen species(RNS)to damage seeds viability.Furthermore,HT increased the S-nitrosylation modification of GSNOR1 protein,and triggered the degradation of GSNOR1,therefore stabilizing ABI5 to suppress seed germination.Consistently,the ABI5 protein abundance was lower in the transgenic line overexpressing GSNOR1,but higher in the gsnor mutant after HT stress.Genetic analysis showed that GSNOR1 affected seeds germination through ABI5 under HT.Taken together,our data reveals a new mechanism by which HT triggers the degradation of GSNOR1,and thus stabilizing ABI5 to suppress seed germination,such mechanism provides the possibility to enhance seed germination tolerance to HT through genetic modification of GNSOR1.

Genetic Variation in Morphophysiological Characters of Seeds of Ceratoides arborescens,a Species Endemic to China

[Objective] The research aimed at investigating the variation in morpho-physiological characteristics of Ceratoides arborescens seeds from Inner Mongolia grassland.[Method] The seeds of six wild C.arborescens populations were collected from Siwangziqi,Xilinhaote,Keshiketengqi and Wengniuteqi in Inner Mongolia grassland;then a total of eight indicators of seed morphology and germination traits were determined,respectively.[Result] There was wide variation in seed traits among different populations of C.arborescens.The variation within the populations was higher than that among the populations,which meant that C.arborescens had a strong adaptability to environment pressure.The seed traits shared a significant positive correlation with each other（r0.931,P0.01）.The three germination-related indices including seed germination rate,germination index,vitality index had significant positive correlations with each other（r 0.836,P0.01）and with seed setting rate（r0.896,P0.01）,but shared weak correlations with morphological traits.There were significant correlations between pappus length and longitude,rainfall,as well as between seed width and temperature,rainfall（r0.801,P0.01）.[Conclusion] The research will provide reference for the seed breeding and collection of germplasm resource.We suggest that collecting seeds from various individuals from each populations will be the preferred strategy to initiate plant improvement program of the species and for gene conservation in seed banks.

The BHLH Transcriptional Factor PIF4 Competes with the R2R3-MYB Transcriptional Factor MYB75 to Fine-Tune Seeds Germination under High Glucose Stress

It is known that the high level of sugar including glucose suppresses seed germination through ABA signal.ABI5 is an essential component to mediate ABA-dependent seed germination inhibition,but underlying mechanism needs more investigation.Previous study demonstrated the PIF4 activated the expression of ABI5 to suppress seed germination in darkness.Here we reported that PIF4 also mediated the seed germination inhibition through ABI5 under high concentration of glucose treatment.Furthermore,we found that PIF4 interacted with PAP1,the central factor to control anthocyanin biosynthesis.Such interaction was confirmed in vitro and in planta.Biochemical and physiological analysis revealed that PAP1 bond the promoter of ABI5 to suppress its expression,thus enhanced seed germination under high concentration of glucose treatment.Specially,PAP1 competed with PIF4 to antagonize the activation of PIF4 on ABI5 expression,thus promoted seed germination under high glucose treatment.Given these,we uncover a novel role for PIF4 and PAP1 in controlling seed germination under high glucose treatment,and reveal their antagonistic mechanism by which coordinates ABI5 expression to control seed germination in response to the glucose signal.

Effect of salinity on seed germination, growth and ion content in dimorphic seeds of Salicornia europaea L. （Chenopodiaceae）

The halophyte Salicornia europaea L. is a widely distributed salt-tolerant plant species that produces numerous dimorphic seeds. We studied germination and recovery in dimorphic seeds of Central Asian S. europaea under various salinity conditions. We also tested the effects of various salts on Na＋ and K＋ accumulation during plant development from germination to anthesis under greenhouse conditions. We found good germination （close to control） of large seeds under NaCl between 0.5 and 2%, Na2SO4 and 2NaCl ＋ KCl ＋ CaCl between 0.5 and 3%, and 2Na2SO4 ＋ K2SO4 -- MgSO4 between 0.5 and 5%. For the small seeds, we found stimulating effects of chloride salts （both pure and mixed） under 0.5-1% concentrations, and sulfate salts under 0.5-3%. Both types of seeds showed high germination recovery potential. Salt tolerance limits of the two seed types during germination and at the later stages of development were very similar （4-5%）. During plant growth the optimal concentrations of mixed chloride and sulfate salts ranged from 0.5 to 2%. The mechanisms of salt tolerance in the two seed types of S. europaea appear to differ, but complement each other, improving overall adaptation of this species to high salinity.

Effects of Space Radiation Mutation on Germination and Growth of Sugarcane Hybrid Seeds

[Objectives] This study was conducted to investigate the mutagenic effects of space radiation mutagenesis on sugarcane hybrid seeds. [Methods] Sugarcane hybrid spikes were subjected to space radiation mutagenesis treatment to identify the effects on the germination, seedling growth and plant growth and development of sugarcane hybrid seeds. [Results] The space radiation treatment had a great impact on the germination and growth of sugarcane hybrid seeds, especially on the germination of sugarcane seeds. The germination number per gram of seeds increased significantly, and the survival rate of seedlings increased significantly. The effect on the growth status was relatively small. The average number of tillers decreased slightly, the percentage of available stems increased, and the plants became higher, which resulted in a greater effect on sugarcane yield. Sugarcane sucrose was increased. The seed germination and late growth status indicated that space radiation promoted the germination and growth of sugarcane hybrid seeds. [Conclusions] This study provides a basis for the space radiation mutagenesis breeding of sugarcane hybrid seeds and the promotion of sugarcane breeding process.

Decontamination of infected plant seeds utilizing atmospheric gliding arc discharge plasma treatment

In agriculture production,plant health is threatened by pathogens parasitic on seeds;hence,it is necessary to disinfect harvested seeds before germination.In this study,a technique of gliding arc plasma treatment was proposed and investigated.The experiment was conducted to treat Astragalus membranaceus(A.membranaceus)seeds that were artificially infected with Fusarium oxysporum(F.oxysporum).The plasma treatment duration varied from 30 s to 270 s.Direct and indirect treatments were compared to evaluate the inactivation efficiency of the F.oxysporum spores on the surface of seeds.The results indicated that the direct treatment behaved significantly better in disinfection than the indirect way.Meanwhile,experiments of the quantitative assessment of seed germination were also conducted,including the germination rate,the germination potential,and the germination index.The results showed that the inactivation efficiency increased as the plasma treatment time was extended.When the treatment time was90 s,the inactivation efficiency reached more than 98%.The plasma treatment of 270 s had a complete devitalization of F.oxysporum spores on the surface of the seeds.After the treatment of 30 s and 90 s,the seed germination parameters improved significantly.This study verified the inactivation efficacy of gliding arc discharge plasma under atmospheric pressure.The technique of gliding arc treatment shows advantages of energy saving and adaptation and has the potential to be utilized in industry.

Conditions and Stimulation for Germination in Glycyrrhiza uralensis Fisch Seeds

Evaluation of seed quality is the key to seed distributing and seeding of Glycyrrhiza uralensis Fisch, as an important species for pharmacy and soil conservation. Here, we study the effects of light and temperature on seed germination and mechanical and chemical scarification on breaking the seed coat. Seeds were collected in 2004, 2005, and 2006, placed in Petri dishes, and incubated at constant temperature 20, 25, and 30℃, and alternating temperature 15-25, 20-30, 15-30, and 20-35℃ under either an 8h photoperiod or total darkness for 28 consecutive days. Different methods were used to break the dormancy owing to hard seededness in this species such as chemical scarification by immersing in concentrated sulphuric acid for 5, 10, 15, 20, 25, 30, 45, and 60 min, in 0.2% KNO3 solution to saturate the seedbed, and prechilling for 7 d at 7℃ and mechanical scarification by cutting. The results showed that alternating temperature at 20-30℃ with 8 h photoperiod and 16 h darkness was optimum for G. uralensis seed germinating in the laboratory. Hard seeds were broken by concentrated sulphuric acid soaking or mechanical scarification by cutting. Germination of seeds harvested in two different years was both promoted by immersing for 30-45 min in concentrated sulphuric acid. KNO3 solution was ineffective for reducing hard seeds. During seed germinating, the first count was on the 7th day and the last count was on the 14th day.

Effect of Natural Brassinolide on Germination of Ailanthus altissima Seeds

The germination capacity of Ailanthus altissima seeds improved after the seeds were soaked with different concentrations of natural brassinolide （NBR）. The germination rate and germination energy of the seeds increased by 17.6% and 18.8%, and the mean germination speed （i.e., germination time） of the seeds was shortened by 1.4 d under the optimal concentration （0.4 mg·L^-1） treatment, compared with the control. After hypocotyls of A.altissima were treated with NBR, the elongation of the hypocotyls increased. Among different concentrations of the NBR, 0.4 mg·L^-1 NBR appeared to be the optimal concentration for the elongation of A.altissima hypocotyls.

Phytotoxic Effects of 4-Chlorophenol and 2,4-Dichlorophenol in the Germination of Seeds of <i>Phaseolus vulgaris</i>and <i>Zea mayz</i>

Soil contaminated with pesticides may reduce plant development due to their toxicity. The aim of this study was to evaluate the influence on the germination of Zea mayz and Phaseolus vulgaris of the two main intermediates of the 2,4-D degradation, which are 4-chlorophenol (4-CP) and 2,4-dichlorophenol (2,4-DCP). Maize and bean seeds were treated with distilled water (control treatment) and increased concentrations of 4-CP and 2,4-DCP (0.006, 0.1, 1.0, 1.5 g·L-1). It was assessed seed germination and calculated various parameters. The parameter most affected by chlorophenols was the index of germination rate, being the P. vulgaris seeds most affected. 2,4-DCP was the compound most toxic for both plants. The germination index was dependent doses for both plant models tested. The results indicate that 4-CP and 2,4-DCP affected the index of germination rate but not influenced in other parameters of germination of Zea mayz and Phaseolus vulgaris. Maize was most tolerance to both chlorophenols in the assessed concentrations. 2,4-DCP was the most toxic of chlorophenols tested.