Deletion of a 1,049 bp sequence from the 5′UTR upstream of the SiHEC3 gene induces a seed non-shattering mutation in sesame

Sesame is a labor intensive crop with limited mechanized harvesting mainly due to the seed shattering(SS)trait.In this study,we performed a genetic analysis of the seed-shattering resistance(SR)trait with a SR sesame mutant 12M07.Unlike the SS type,the parenchyma cells in the abscission zone of the 12M07 mutant are arranged loosely but adhere to the seed coat.Inheritance analysis of six generations derived from 12M07(SR)×Xiangcheng Dazibai(SS)showed that the SR trait is recessive and controlled by a single gene pair.Association mapping of the F2population with 888,619 variants(single-nucleotide polymorphisms(SNPs)and insertion-deletion(InDels))and 31,884 structural variations(SVs)determined that only SV12002 in the 5′upstream region of gene Sindi0765000(named SiHEC3)in Chr.3 was significantly associated with the SR trait.SiHEC3 encodes the bHLH transcription factor.A 1,049 bp deletion occurred in the 5′UTR of Sihec3 in 12M07.SiHEC3 is mainly expressed in developing placental tissues,with the expression peaking in capsules at 45 days after pollination.A dual-luciferase reporter assay in tobacco confirmed that the promoter activity of Sihec3 was reduced because of the deletion of the 1,049 bp promoter sequence.Protein–protein interaction network analysis showed that HEC3 is co-expressed with nine key proteins,such as SHATTERPROOF1(SHP1)and SEEDSTICK(STK)which participate in the secondary wall biosynthesis of the abscission layer in plants.The findings of this study show the important function of Sihec3corresponding with the SR trait and supply the genetic information for breeding new varieties that are amenable to mechanized harvesting in sesame and other crops.

Dormancy of Amaranthus retroflexus L. Seeds and Physiological Response Seedlings to Acifluorfen Sodium

Amaranthus retroflexus L. is a serious and widespread malignant weed in soybean fields in Heilongjiang Province. Exploring the dormancy characteristics of A. retroflexus L. seeds and the physiological response of its seedlings to acifluorfen sodium can provide a basis for further researches on its resistance mechanism. Using newly harvested and stored A. retroflexus L. seeds for one year as experimental materials, the effects of different concentrations of HCl, NaOH, water temperature, gibberellic acid(GA) and polyethylene glycol(PEG) on the dormancy and germination of A. retroflexus L. seeds were studied. The sensitivity of A. retroflexus L.to acifluorfen sodium was determined using bioassay. The effects on leaf chlorophyll content and target enzyme activity were studied at a normal dosage of 360 g a.i. hm^(-2) and a doubling dosage of 720 g a.i. hm^(-2) of acifluorfen sodium. Newly harvested seeds exhibiting dormancy were soaked in water of various temperatures and in different concentrations of NaOH and HCl, which were ineffective in breaking the seed dormancy. GA could break seed dormancy, and the highest seed germination rate reached 93.33% when they were soaked at 3 000 mg·L^(-1) for 72 h and 4 000 mg·L^(-1) for 48 h. The drought stress was simulated with a 15%-25% polyethylene glycol solution, which had no significant effect on the seed germination rate. The GR_(50) value of acifluorfen sodium for A. retroflexus L. was 705.7 g a.i. hm^(-2), which was 1.96 times the recommended dose in the field. After the application of different doses of acifluorfen sodium, the chlorophyll content of A. retroflexus L. reached its minimum value 3 days after treatment(DAT), and then gradually increased. The activity of the target enzyme protoporphyrinogen oxidase(PPO) reached the highest value at 7 DAT under different dosages, and gradually returned to normal levels thereafter. Soaking with gibberellin was an effective method to break seed dormancy. A. retroflexus L. seeds had certain drought resistance during the germination process. A. retroflexus L. was not sensitive to acifluorfen sodium and acifluorfen sodium ether, and could not effectively inhibit the PPO activity, indicating that A. retroflexus L. had target resistance to acifluorfen sodium.

Development of a KASP Marker on Chromosome A05 for Seed Oil Content of Brassica napus

In order to identify the molecular markers that can be widely used in the breeding of Brassica napus L.varieties with high seed oil content under different genetic backgrounds,we developed a Kompetitive Allele Specific PCR(KASP)marker for seed oil content on the basis of the results from available studies.The verification in the F_(2) population showed that the marker was closely linked to the quantitative trait locus(QTL)for oil content on chromosome A05.The findings helped to breed the‘Fengyou’varieties with high seed oil content in the middle reaches of the Yangtze River.

Variations in seed germination ofHippophae salicifolia with different presoaking treatments

Mature seeds of H. salicifolia, collected from five provenances （i.e. Hanuman Chatti, Helang, Lata, Rambara and Janggal Chatti） in west Himalaya, India, were treated with stratification （at 4℃ for 15, 30 and 60 days） and in different concentrations of GA3 （5, 10, 20 mM）, KNO3 （50, 100, 200 mM） and Thiourea （50, 100, 200 mM） solution to determine the variations in seed germination. Results reveal that the germination rates of seeds from different provenances under different pre-sowing treatments are significantly increased compared to those in control （24%-30%）. The seeds treated with Thiourea （100 mM） have highest germination rate （76%-83% for different seed sources）, followed by those （63%-71% for different seed sources） pretreated with stratification （4℃, 30 days）. GA3 treatment significantly shortens the mean germination time （MGT） and improves seed germination percentage. Considering the practical applicability and cost effectiveness, thiourea （100 mM） and stratification （at 4℃） treatments for seed germination are recommended for mass multiplication through seeds of H. saliciifolia in village/forest nurseries of the west Himalaya, India.

Short-term responses to salinity of seabuckthorn (Hippophae rhamnoides L.) seedlings in the extremely cold and saline Qinghai region of China

The investigation of the mechanisms of plant adaptation to stressor action is one of the leading directions of current biological studies. To understand the mechanism of salt tolerance of seabuckthom （Hippophae rhamnoides L.） and identify its ability to cope with the salinity effect in the saline and extremely cold region of Qinghai, China, a test was conducted with two-year-old seedlings subjected to 0, 200, 400 and 600 mmol·L^-1 NaClsolutions for 30 d. The results show that with an increase in salinity, the biomass of H. rhamnoides seedlings clearly decreased. Leaf water potential （ψw） and relative water content （RWC） were significantly reduced under salinity, with severe water shortages appearing in leaves. At the same time, the total chlorophyll content declined markedly. When salinity increased and stress time prolonged, the net CO2 assimilation rate （A） significantly declined. Intercellular CO2 concentration （Ci） declined at first and was then followed by an increase over the stress time. We conclude that H. rhamnoides grown in the extremely cold and saline region of Qinghai has a certain resistance to salt, which can be planted at appropriate salinity levels.

Advances in Physiological-biochemical and Genetic Mechanisms of Seed Aging in Rice(Oryza sativa L.)

Reasons causing or accelerating seed aging are mainly damage of mem- branes, DNA and proteins, decline of protein synthesis capacity and excessive ac- cumulation of reactive oxygen species. With the application of natural aging or artifi- cial aging methods, it was reported that quantitative trait loci （QTLs） of seed stora- bility in rice were widely distributed on the chromosomes except the 10th chromo- some. In this paper, we reviewed the progresses in the research on physiological- biochemical and genetic mechanisms of seed aging, and analyzed the existing problems and developing prospect in molecular breeding of rice with improved seed storability, in order to provide reference for the basic research and genetic improve- ment of rice seed storabUity.

Reproduction of the Three-line Genic Male Sterile Line Parent Mian 7MB-1 （Brasscia Napus L.） and Seed Production of F1 Based on Somatic Tissue Culture

[Objective] The aim was to study the reproduction of the three-line genic male sterile （GMS） lineparent Mian7MB-1 （B. NapusL.） and the seed production of F1 through somatic tissue culture. [Methed] Through hybridization, a new breeding material Mian 7MB-1 in three-line genic temporary maintainer line propagated by tissue culture was used to improve the sterile plant rate of rapeseed in dual-purpose recessive GMS line, such as Mian 7AB type, S45AB type, and etc. And then the variety comparative test was performed. [Result] In order to avoid some fertility restoration phenomena occurring during the process of self-reproduction, Mian 7AB was propagated in bulk with somatic tissue culture of temporary maintainer line plant stem. The propagated temporary maintainer line seedlings were applied to the breeding and seed production of net room male sterile line parent, promoting the sterile plant rate of the male sterile line parent to 91.7% -93.5%. The male sterile line parents per hectare were enough for the seed production of hybrid F1 in 7 500 -15 000 hm^2. [ Conclusion ] Compared with the original dual-purpose GMS line, the seed production ultilizing male sterile line with high sterile plant rate greatly reduced the labor, significantly improved the seed yield, ensuring the seed quality and forming a perfect breeding and seed production system.

Effects of Cadmium Stress on Seed Germination and Seedling Growth of Brassica napus L.

To investigate the effects of the heavy metal cadmium （Cd） on seed ger- mination and seedling growth of Brassica napus L., 25 B. napus lines with different genetic background were treated with four concentrations （0, 1, 5 and 10 mg/L） of Cd, and then, their germination potential, germination rate, seedling length, main root length, seedling fresh weight, and seedling dry weight were measured. The re- sults showed that the response of the B. napus lines to Cd stress was different. With the increase of Cd concentration, the relative germination potential and relative seedling rate increased first and then decreased, and reached the maximum levels when the Cd concentration was 5 rag/L. The inhibitory effects of Cd stress on seedling growth indices in a decreasing order were root length〉seedling length〉 seedling fresh weight〉seedling water content. There were extremely significant differ- ences in the relative root length, relative seedling length, relative seedling fresh weight and relative seedling water content among the B. napus lines treated by dif- ferent concentrations of Cd, while there were no significant differences in the rela- tive germination potential and relative seedling rate among the lines, except that the relative seedling rate was significantly different among the lines treated by 5 mg/L Cd. The results suggested that seedling growth was more sensitive than seed ger- mination to heavy metal stress.

Effects of Salt Stress on Seed Germination and Seedling Growth in Raphanus sativus L.

[Objective] This study aimed to investigate the effects of salt stress on seed germination and seedling growth in radish（Raphanus sativus L.）.[Method] First,effects of soaking seeds with clear water and different concentrations of saline solution were compared; then,different concentrations of salt solution （0.2％-1.0％） was used to treat radish during the whole germination period.[Result] soaking seeds with clear water can improve the germination indicators of radish seeds exposed to high salt stress; low salt concentration（0.2％,0.4％） had little effect on the growth indicators of radish seedlings,and 0.2％ salt concentration can even promote the growth of radish seedlings,however,the high salinity had a bad inhibition on seedling growth,the higher the salt concentration,the stronger the inhibition.[Conclusion] According to the comprehensive comparison,the order of the six varieties in salt-tolerance was as follows：Xinxiayuanbai＞W1029,Xiayuanbai ＞ Nanyangbailizhi,Chunyidianhong＞ Sijihong 3.

Study on Seed Vigor of Rice Photo-ThermoSensitive Genic Male Sterile(PTGMS) Line with Low Temperature Storage

To explore the change rule of seed vigor and seed application technology of rice PTGMS lines with low temperature storage,Zhun S,Y58S,Feng S and Guangzhan 63-4S stored in the seed bank with the temperature of 8-10 ℃ and with the relative humidity of 55% for different years were used as the materials to study the properties of seed vigor,growth and development in the field, etc. The re- sults showed that the germination potential, germination rate, germination index and vigor index of rice PTGMS lines seeds showed downward trends as the time of low temperature storage prolonged,and all of the germination rates of the seeds within five years were over 60%. The conductivity in the soaked solution of seeds was significantly increased with the prolonged low temperature storage time. Seedling rate,root length,white root number,base stem width and seedling dry weight of seeds showed downward trends as the time of low temperature storage prolonged. There was no significant difference between two integral seedling qualities of seeds within 4-6 years in low temperature storage. Compared with the germination rate of low temperature storage seeds soaking for 6,12 and 24 h,the appropriate soak time of long time low temperature storage or low vigor seeds was 12 h. A serious natural aging phenomenon occurred in low temperature storage seeds after out of the storage. The germination rate of seeds placed under natural condition for 50 d decreased obviously. The duration from seeding to heading of Zhun S,Y58S and Feng S shortened as the time of low temperature storage prolonged, while the vari- ation of panicle duration of all PTGMS lines was not obvious. In conclusion, rice PTGMS line seeds with low temperature storage in 5 years could still be used nor- mally. However.seeds after low temperature storage should be sowed as soon as possible, and the soaking time of low vigor seeds should be shortened,and the possibility that the decreased duration from seeding to heading should also be con- sidered.

Effects of Nitrogen Fertilizer Management on Biological Traits and Physiological Indexes at Seedling Stage and Yield of Directly-sown Rapeseed(Brassica napus L.)

Field experiments were carried out to study the effects of different nitroge- napplication rates and application times on biological traits and physiological indexes of directly-sown rapeseed （Brassica napus L.） at the seedling stage and investigate the relationship between these biological traits or physiological indexes at the seedling stage and yield, so as to provide scientific theoretical support for high yield and efficient fertilization management in production of winter rapeseed. Field trials were conducted in Chengdu plain of Sichuan Province under rice-rapeseed rotation system during the period of 2011-2012. The nitrogen application rate trial consisted of five nitrogen levels （0, 90, 180, 270 and 360 kg/hm2） and the nitrogen application time trial included NTl（single application as base fertilized）, NT2 （bottom application ＋one time of topdressing at seedling stage） and NT3 （bottom application＋two times of topdressing at seedling stage） under the same nitrogen rate （225 kg/hm2）. The results indicated that compared with no nitrogen application （NO） treatment, the in- crease of nitrogen fertilizer is beneficial to the increase of biological traits including plant height, green leaf number, leaf area index and dry weight of rapeseed at the seedling stage, the improvement of physiological indexes including total nitrogen content, chlorophyll content and soluble protein content of functional leaves, and the reduction of soluble sugar content. Nitrogen rate was linearly correlated with various biological traits at the seedling stage and physiological indexes including total nitro- gen content, chlorophyll content and soluble sugar content in functional leaves over- a/I, but in parabolic correlation with soluble protein content. Under the same nitrogen rate, NT2 treatment exhibited biological traits remarkably or significantly higher than NT1 treatment and NT3 treatment. The nitrogen application times were linearly cor- related with the physiological indexes of functional leaves at the seedling stage. The various biological traits and physiological index of functional leaves at the seedlings stage were in quadratic function parabolic correlation with seed yield, and the corre- lation was significant （P〈0.05）. Therefore, under the rice-rapeseed rotation system in Chengdu plain, the economic rational nitrogen rate is 180-225 kg/hm2, and the mode of bottom application ＋ one time of topdressing （NT2） is suitable.

Allelopathic effects of extracts from Solidago canadensis L. against seed germination and seedling growth of some plants

To investigate the potential role of allelopathy in plant interference and in the successful invasion of alien species Solidago canadensis, aqueous and ethanolic extracts from rhizomes, stems and leaves of S. canadensis were prepared and used as treatment solutions to assess their effects on seed germination and seedling growth in four target species, mulberry （Morus alba）; morning glory （Pharbitis nil）, wheat （Triticum aestivum） and rape （Brassiea campestris）. Reduction and/or growth in germination and growth of the target plant species in the presence of both aqueous and ethanolic extracts at different concentrations indicated that the responses were species-specific and concentration-dependent. Generally, ethanolic extracts （especially from leaves） imposed stronger effects on both seed germination and seedling growth. Extracts with lower concentration at 0.001 g/ml dw could stimulate the seedling growth of rape and morning glory, whereas extracts at any given concentrations have inhibitory effects on wheat and mulberry. It is suggested that the aqueous and ethanolic extracts of all the three parts of S. canadensis have significant allelopathic effects. Although both inhibition and stimulation occurred in the germination and growth of the target species, extracts with higher concentrations definitely inhibit seed germination and seedling growth of all target plants. We suggest that allelopathy plays a more important role than other mechanisms do in the out-competition ofS. canadensis over other plants, and make it invasive in new habitats.

Mechanical Harvesting Effects on Seed Yield Loss,Quality Traits and Profitability of Winter Oilseed Rape (Brassica napus L.)

China is one of the most important rapeseed producing countries in the world. Effective mechanical harvesting time for decreasing harvesting loss of winter oilseed rape has been becoming a critical factor. An elite cultivar Zhongshuang 11 （Brassica napus L.） was employed in two rounds of field experiments from 2009 to 2011. Seeds were sown with machine, three combine harvesting times namely combine harvesting A, B, and C （CHA, CHB, and CHC） were designed and manual harvesting （MH） as control was performed at maturity. The harvesting treatments were determined according to color of pod and seed in the field. Seed yield loss and quality in different treatments were evaluated. Results showed that both seed yields and harvesting losses in 2009-2010 were higher than that in 2010-2011, whereas seed oil contents in 2010-2011 were higher than that in 2009-2010. The highest yield appeared in CHB, which was significantly higher than that in MH. Furthermore, harvesting loss in CHB were 50% that in MH. Seed oil content and chlorophyll exhibited no obvious difference between CHB and MH. Economic profit analysis demonstrated that mechanical sowing/combine harvesting （MS/CH） showed an input/output ratio of 1：1.6, and it was 1：1.2 in mechanical sowing/manual harvesting （MS/MH）. Labor-cost accounted for more than 70% of the total cost in MS/MH, which led to low profitability to a great extent. Our results suggested that CHB was the optimum harvesting time for winter oilseed rape along the Yangtze River.

Comparative QTL analysis of maize seed artificial aging between an immortalized F_2 population and its corresponding RILs

Seed aging decreases the quality and vigor of crop seeds,thereby causing substantial agricultural and economic losses in crops.To identify genetic differences in seed aging between homozygotes and heterozygotes in maize,the seeds of a set of recombinant inbred lines(RILs) and an immortalized F_2(IF_2) population were subjected to artificial aging treatments for 0,2,3,and 4 days under 45℃ and 85%relative humidity and seed vigor was then evaluated in a field experiment.Seed vigor of all entries tested decreased sharply with longer aging treatment and seed vigor decreased more slowly in heterozygotes than in homozygotes.Forty-nine QTL were detected for four measured seed vigor traits in the RIL(28QTL) and IF_2(21 QTL) populations.Only one QTL,qGP5,was detected in both populations,indicating that the genes involved in anti-aging mechanisms differed between inbred lines and hybrids.Several QTL were identified to be responsible for multiple seed vigor traits simultaneously in the RIL and IF_2 populations under artificial aging conditions.These QTL may include major genes for seed vigor or seed aging.QTL qVI4 b and qGE3 a detected in the RIL population coincided with genes ZmLOX1 and ZmPLD1 in the same respective chromosomal regions.These QTL would be useful for screening for anti-aging genes in maize breeding.

Nutrient deficiency limits population development,yield formation,and nutrient uptake of direct sown winter oilseed rape

Direct-sowing establishment method has great significance in improving winter oilseed rape（Brassica napus L.） production and guaranteeing edible oil security in China. However, nutrient responses on direct sown winter oilseed rape（DOR） performance and population development dynamic are still not well understood. Therefore, five on-farm experiments were conducted in the reaches of the Yangtze River（RYR） to determine the effects of nitrogen（N）, phosphorus（P）, and potassium（K） deficiencies on population density, dry matter production, nutrient uptake, seed yield, and yield components of DOR plants. Four fertilization treatments included the balanced NPK application treatment（NPK, 180 kg N, 39.3 kg P, 100 kg K, and 1.8 kg borax ha–1） and three nutrient deficiency treatments based on the NPK treatment, i.e., –N, –P, and –K. The results indicated that DOR population density declined gradually throughout the growing season, especially at over-wintering and pod-development stages. Nutrient deficiency decreased nutrient concentration in DOR plants, limited dry matter production and nutrient uptake, and thereby exacerbated density reduction during plants growth. The poor individual growth and reduced population density together decreased seed yield in the nutrient deficiency treatment. Averaged across all the experiments, seed yield reduced 61% by N deficiency, 38.3% by P deficiency, and 14.4% by K deficiency. The negative effects of nutrient deficiency on DOR performances followed the order of –N–P–K, and the effects were various among different nutrient deficiencies. Although N deficiency improved DOR emergence, but it seriously limited dry matter production and nutrient uptake, which in turn led to substantial plants death and therefore resulted in a very low harvested density. The P deficiency significantly reduced initial density, limited plants growth, and exacerbated density reduction. The K deficiency mainly decreased individual growth and yield, but did not affect density dynamic. Our results highlighted the importance of balanced NPK application in DOR production, suggesting that management strategy of these nutrients should be comprehensively considered with an aim to build an appropriate population structure with balanced plant density and individual growth.

Identification of a novel seed size associated locus SW9-1 in soybean

Seed size is one of the vital traits determining seed appearance, quality, and yield. Untangling the genetic mechanisms regulating soybean 100-seed weight (100-SW), seed length and seed width across environments may provide a theoretical basis for improving seed yield. However, there are few reports related to QTL mapping of 100-SW across multiple ecological regions. In this study, 21 loci associated with seed size traits were identified using a genome-wide association of 5361 single nucleotide polymorphisms (SNPs) across three ecoregions in China, which could explain 8.12%–14.25% of the phenotypic variance respectively. A new locus, named as SW9-1 on chromosome 9 that explained 10.05%–10.93% of the seed weight variance was found significantly related to seed size traits, and was not previously reported. The selection effect analysis showed that SW9-1 locus has a relatively high phenotypic effect (13.67) on 100-SW, with a greater contribution by the accessions with bigger seeds (3.69) than the accessions with small seeds (1.66). Increases in seed weight were accompanied by increases in the frequency of SW9-1T allele, with >90% of the bred varieties with a 100-SW >30 g carrying SW9-1T. Analysis of SW9-1 allelic variation in additional soybean accessions showed that SW9-1T allele accounting for 13.83% of the wild accessions, while in 46.55% and 51.57% of the landraces and bred accessions, respectively, this results indicating that the SW9-1 locus has been subjected to artificial selection during the early stages of soybean breeding, especially the utilization of SW9-1T in edamame for big seed. These results suggest that SW9-1 is a novel and reliable locus associated with seed size traits, and might have an important implication for increasing soybean seed weight in molecular design breeding. Cloning this locus in future may provide new insights into the genetic mechanisms underlying soybean seed size traits.

Effects of Total Flavonoids ofHippophae RhamnoidesL.on Intracellular Free Calciumin Cultured Vascular Smooth Muscle Cells of Spontaneously Hypertensive Rats and Wistar-Kyoto Rats

To explore the effects of total flavonoids of Hippophae rhamnoides L. （TFH） quercetin （Que） and isorhamnetin （Isor） on the intracellular free calcium （[Ca^2＋]） in vascular smooth muscle cells （VSMC） of spontaneously hypertensive rats （SHR） and Wistar-Kyoto rats （WKY）. Metheds： Fluo 3-acetoxymethylester（Fluo-3/AM） was used to observe the effects of TFH （100mg/L） and its essential monomers, namely Que （10^-4mol/L） and Isor （10^-4mol/L） on changes of [Ca^2＋]1 in cultured SHR and WKY VSMC （abbr. to Ca-SHR ＆ Ca-WKY） following exposure to high K^＋, norepinephrine （NE） and angiotensin Ⅱ （AngⅡ）, and to compare with the effects of verapamil （Ver）. Results： （1） TFH, Que and Isor had inhibitory effects on resting Ca-SHR （P〈0.05）, but had no significant effects on Ca-WKY （P〉0.05）. （2） High K^＋ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）; TFH, Que and Isor could inhibit the elevation of [Ca^2＋]1 induced by high K^＋ -depolarization, with the effects similar to that of Ver, and the effect on Ca-SHR was more significant than that on Ca-WKY （P〈0.05）. （3） NE and Ang Ⅱ could increase Ca-SHR more significantly than Ca-WKY （P〈0.05）, TFH, Que and Isor had remarkably inhibitory effect on the elevation of Ca-SHR and Ca-WKY induced by NE or Ang Ⅱ. （4） In the absence of extracellular Ca^2＋ , TFH, Que and Isor also had certain inhibitory effect on Ca-SHR and Ca-WKY induced by NE, and the effect on the former was more significant than that on the latter（P〈0.05）. Ceaclusiea： TFH, Que and Isor might decrease the levels of [Ca^2＋], in VSMCs by blocking both voltage-dependent calcium channels （VDC） and receptoroperated calcium channels （ROC） in physiological or pathological state, which may be one of the important mechanisms of their hypotensive and protective effects on target organs in patients with hypertension.

Molecular identification of sex in Hippophae rhamnoides L. using isozyme and RAPD markers

In many dioecious plants, gender affects economic value, breeding schemes and opportunities for commercial harvests. Hippophae rhamnoides L. is a dioecious plant species in which female genotypes are commercially preferred over male genotypes. Its berries have rich medicinal, nutritional and pharmaceutical properties because of their large amounts of vitamins, essential oils, proteins, fatty acids, free amino acids and flavanoids. Primary limitation for breeding H. rhamnoides L. is its dioecious nature, since gender cannot be identified by traditional methods. Therefore, some reliable and quick methods need to be developed. This commu- nication deals with the development of isozyme and RAPD markers for early sex identification in this dioecious tree. The isozyme analysis was conducted with four enzyme systems, viz. peroxidase, esterase, malate dehydrogenase and catalase. The peroxidase enzyme system produced a female specific sex marker, which successfully differentiated between the staminate and pistillate geno- types ofH. rhamnoides L. Thirty five random decamer primers were used in our study and one male sex linked marker was identified. OPD-20 （5＇-ACTTCGCCAC-3＇） displayed a band at 911 bp that expressed polymorphism between male and female genotypes. The staminate and pistillate genotypes could be distinguished using RAPD marker OPD-209n. These results revealed the immense poten- tial of peroxidase isozyme patterns and RAPD as genetic markers for sex identification in H. rhamnoides L.

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.

Interaction between seed size and NaCl on germination and early seedling growth of some Turkish cultivars of chickpea(Cicer arietinum L.)

Chickpea is an important food legume crop of Turkey and is largely grown for human consumption on low moisture or salt-affected soils. The objective of the study was to find the effects of NaCl stress at electrical conductivities of 4.5, 8.6, 12.7 and 16.3 dS/m and seed sizes （7, 8 and 9 mm） on germination and early seedling growth of three popular chickpea cultivars （AKN-97, Gokce and Uzunlu-99）. Mean frequency of germination, germination time, germination index, root length, shoot length and seedling fresh weight showed seed size-dependent responses ofcultivars to salt stress. In general, small seeds germinated and grew more rapidly compared to medium and large seeds of the same cultivars against all levels of salt stress, with the best results in cultivar Uzunlu-99. No effect of NaCl treatments was observed on frequency of germination; however, a drastic decrease in early seedling growth was recorded at increased NaC1 concentrations. Regression analysis results showed a significantly positive relationship （P〈0.01） between seed size and mean germination time, whereas a significantly negative relationship was recorded between seed size and germination index, root length, shoot length. Moreover, linear regression values apparently confirmed that increased seed size in each cultivar affected decreased germination index, root and shoot lengths with enhanced mean germination time. Thus, it was concluded that the use of small seeds could considerably reduce the production costs of chickpea in salt-affected soils.