Genetics of drought tolerance during seed germination in tomato: inheritance and QTL mapping.

A BC1 population (N = 1000) of an F1 hybrid between a stress-sensitive Lycopersicon esculentum breeding line (NC84173; maternal and recurrent parent) and a germination stress-tolerant Lycopersicon pimpinellifolium accession (LA722) was evaluated for seed germination rate under drought stress (DS) (14% w/v polyethyleneglycol-8000, water potential approximately -680 kPa), and the most rapidly germinating seeds (first 3% to germinate) were selected. The 30 selected BC1 seedlings were grown to maturity and self pollinated to produce BC1S1 progeny seeds. Twenty of the 30 selected BC1S1 progeny families were evaluated for germination rate under DS and their average performance was compared with that of a "nonselected" BC1S1 population of the same cross. Results indicated that selection for rapid germination under DS significantly improved progeny germination rate under DS (selection gain = 19.6%), suggesting a realized heritability of 0.47 for rate of germination under DS in this population. The 30 selected BC1 plants were subjected to restriction fragment length polymorphism (RFLP) analysis, and marker allele frequencies for 119 RFLP markers which spanned 1153 cM of the 12 tomato chromosomes were determined. A distributional extreme marker analysis, which measures statistical differences in marker allele frequencies between a selected and a nonselected population, detected four quantitative trait loci (QTLs) for rate of germination under DS in this population. Of these, two QTLs, located on chromosomes 1 and 9, were contributed by the L. pimpinellifolium donor parent and had larger effects than the other two QTLs, located on chromosomes 8 and 12, which were contributed by the L. esculentum recurrent parent. A few BC1S1 families were identified with all or most of the identified QTLs and with germination rates comparable with that of LA722. These families should be useful for the development of germination drought-tolerant tomato lines using marker-assisted selection (MAS). The overall results indicate that drought tolerance during seed germination in tomato is genetically controlled and potentially could be improved by directional phenotypic selection or MAS.     

The influence of temperature during seed development on the germination characteristics of millet seeds

Abstract. The cardinal temperatures, rate of germination and final percentage germination of pearl millet seeds were measured for seeds raised in greenhouses maintained at mean air temperatures of 19, 22, 25, 28 and 31°C. The results showed that cardinal temperatures for germination are unaffected by the temperature during seed development and growth. However, the conditions during seed growth did affect seed size and, subsequently, germination rate and seed viability.     

The effect of light and exogenous gibberellic acid on respiration pathways during germination of tomato seeds

Tomato (Lycopersicon esculenlum Mill. cv. Taiwan Red) seeds are typical lightsensitive seeds where light requirement can be substituted by gibberellic acid (GA₃). During the initial stage of germination, the hexose monophosphate pathway (HMP) in seeds incubated for 24 h under white light or 12 h in 0.3 mW GA₃ solution acidified with 1 M HCI for 1 h in the dark (HCl→GA₃) was much greater than in seeds incubated for 24 h in the dark. The results were obtained from measurements of the respiration rate by man metric method of Warburg with or without iodoacetic acid, and from activities of glucose-6-phosphate dehydrogenase (EC 1.1.1.49) and aldolase (EC 4.1.2.13). Although the HMP tended to decrease at the later stage (72 h under white light or in the dark, or 36 h in HCl→GA₃), it remained greater in light- than in dark-incubated seeds. CN-resistant respiration increased from 27–30% to 57–64% of the total respiration in seeds incubated under white light or in HCI→GA₃, while respiration of non-germinating, dark-incubated seeds remained zero. Benzohydrox–amic acid (BHAM), an inhibitor of the alternative pathway, inhibited both respiration and seed germination. It is concluded that the enhancement of HMP and the CN–resistant pathways are both controlled by P_fr, but there is no direct connection between them.     

Developmental expression and perturbation of arabinogalactan-proteins during seed germination and seedling growth in tomato

Arabinogalactan-proteins (AGPs) are a family of highly glycosylated hydroxyproline-rich glycoproteins present throughout the plant kingdom. A synthetic chemical reagent, ( β - d -Gal)₃ Yariv reagent, specifically binds AGPs and can be used for histochemical staining, isolating and probing the function of AGPs. Here, the role of AGPs in tomato ( Lycopersicon esculentum Mill. cv. UC82B) seed germination and seedling growth was examined by following expression of AGPs during these events and by treatment with ( β - d -Gal)₃ Yariv to perturb AGP function. AGP expression changed during germination and seedling development both quantitatively and qualitatively as revealed by analysis of total AGP content, crossed electrophoresis patterns, RNA blots using LeAGP-1 probe, and western blots with LeAGP-1, JIM13, and MAC207 antibodies. ( β - d -Gal)₃ Yariv treatment of seeds and developing seedlings did not affect percent seed germination, but markedly inhibited seedling growth in roots and to a lesser degree in shoots. Root growth inhibition encompassed reductions in overall root length, epidermal root cell elongation, root cell numbers and root hair formation. This growth inhibition was reversible following removal of ( β - d -Gal)₃ Yariv. In a related experiment, water uptake by tomato seedlings was greatly inhibited by ( β - d -Gal)₃ Yariv treatment. Based on these experiments, AGPs are clearly associated with tomato seedling development and likely to function in root growth, more specifically in cell elongation, cell proliferation, root hair formation and water uptake.     

Hormonal regulation of tomato seed germination at a supraoptimal temperature

Germination of tomato cv. New Yorker seed is inhibited at 35°C. This thermoinhibition was partially counteracted by application of GA₄₊₇ alone, the compound applied in combination with ACC or ethephon markedly enhancing the process. The latter compound alone was not able to induce germination at 35 °C. Thermoinhibition of seeds at 35 °C was also counteracted by fluridone, an inhibitor of ABA biosynthesis. At 25 °C, an optimal temperature, ABA inhibited germination of New Yorker seeds. Although another known growth inhibitor MeJA, when applied at an optimal temperature (25 °C), had also a slightly inhibitory effect on germination of those seeds and clearly delayed the process, inhibitors of its biosynthetic pathway (ibuprofen, indoprofen, antypiryne and salicylic acid) did not remove thermoinhibition at 35 °C. An increase in endo-β-mannanase activity after 24 hours of incubation at 35 °C was observed in the seeds incubated in the presence of gibberellins, ACC, ethephon, fluridone used alone and in combinations, but it was not clearly correlated with the effects of these compounds on alleviation of seed germination. However, fluridone present in the same incubation medium at 35 °C with ABA was able to counteract the inhibitory effect of ABA on endo-β-mannanase activity. The results of our study suggest that gibberellins, ethylene (produced from ACC or ethephon) and ABA, but not jasmonates, regulate tomato seed germination at supraoptimal temperatures. Alleviation of thermoinhibition of New Yorker seed germination by plant growth regulators and fluridone is partially associated with their controlling endo-β-mannanase activity.     

Phytochrome and seed germination. V. Changes of phytochrome content during the germination of cucumber seeds

Cucumber seeds are light-sensitive, dark-germinating seeds. Inhibition of germination can be induced by prolonged exposure to continuous or intermittent FR. The dark germination process and the response to FR are phytochrome controlled. Phytochrome can be detected in these seeds by differential spectrophotometry in vivo. Spectrophotometrically measurable phytochrome increases during dark germination. The rate of increase is temperature dependent. Light treatments which are inhibitory for germination result in phytochrome contents lower than those of the seeds germinating in darkness. Treatments which restore germination also restore phytochrome formation.     

Investigation of the relationship between seed leachate conductivity and the germination of Brassica seed

Large differences in the leachate conductivities of single seeds after 24 h soaking were observed in five Brussels sprout cultivars. Within each cultivar non-germinable seeds tended to have higher conductivities than germinable seeds, but the conductivity above which seeds failed to germinate varied between cultivars. No conductivity reading or partition value could therefore be considered as an absolute boundary between germinable and non-germinable seeds to allow the prediction of seed germination for all seed lots of the same species. However, the mean conductivity for each sample was significantly correlated with germination (r = 0.83, P < 0.05) and was clearly higher for samples with lower germinations. Differences in the partition values for the five cultivars may have been attributable to the natural age of the seeds. Seeds of Asmer Aries artificially aged rapidly at 20% moisture content for up to 48 h at 45° C showed increasing leachate conductivity with increased time of ageing (r = 0.97, P < 0.01) and decreased germination (r = - 0.88, P < 0.05). Again however the values for germinable and non-germinable seeds of different artificial ages varied. Leakage from seeds was limited by the testa in the first 6 h of imbibition. Solute leakage from embryos of the five cultivars was therefore greater than those for intact seeds but a clear separation of germinable and non-germinable seeds was still not possible from the conductivity readings. The implications of the observed increases in conductivity with increased seed age and decreased germination for the development of a rapid vigour test for small seeded vegetables in conjunction with controlled deterioration are discussed.     

The influence of abscisic acid on different polysomal populations in embryonal tissue during pea seeds germination

The influence of abscisic acid (ABA) on the processes of formation of different polysomal populations, their structures and stability in embryonal tissue during pea seeds germination was studied. The contents of total ribosomal fraction increased in all samples up to 72 h of germination and then decreased. The contents of polysomal population (FP, MBP, CBP and CMBP) extracted from the embryonal tissue after 72 hrs of germination of pea seeds were then quantified. It turned out that in examined tissue of control sample, fraction of free polysomes (FP) was the most abounded. This population of polysomes in sprouts decreased after ABA treatment. FP content decreased even more when the higher ABA concentration was applied during germination. Similar changes were observed in the fraction of membrane-bound polysomes (MBP). Quite different tendencies were found, however, in forming population of the cytoskeleton-membrane-bound polysomes (CMBP). The CMBP population content in embryonal tissue increased in a dosage dependent manner with increasing concentration of ABA applied during seed germination. This indicates the important role of CMBP fraction in synthesis of specific proteins in embryos in the time when processes of seeds germination are retarded by ABA. In the final part we examined the stability of polysomes isolated from sprouts of germinating seeds in water and sprouts isolated from seeds treated with ABA (100 μM) during germination. Total polysomes isolated from embryonal tissue of germinating seeds treated with ABA showed much higher resistance to exogenous ribonuclease digestion than total polysomes of control sample. The obtained results suggest that ABA influence on different polysomal population formation also controls their stability.     

Expression of an expansin is associated with endosperm weakening during tomato seed germination

Expansins are extracellular proteins that facilitate cell wall extension, possibly by disrupting hydrogen bonding between hemicellulosic wall components and cellulose microfibrils. In addition, some expansins are expressed in non-growing tissues such as ripening fruits, where they may contribute to cell wall disassembly associated with tissue softening. We have identified at least three expansin genes that are expressed in tomato (Lycopersicon esculentum Mill.) seeds during germination. Among these, LeEXP4 mRNA is specifically localized to the micropylar endosperm cap region, suggesting that the protein might contribute to tissue weakening that is required for radicle emergence. In gibberellin (GA)-deficient (gib-1) mutant seeds, which germinate only in the presence of exogenous GA, GA induces the expression of LeEXP4 within 12 hours of imbibition. When gib-1 seeds were imbibed in GA solution combined with 100 microM abscisic acid, the expression of LeEXP4 was not reduced, although radicle emergence was inhibited. In wild-type seeds, LeEXP4 mRNA accumulation was blocked by far-red light and decreased by low water potential but was not affected by abscisic acid. The presence of LeEXP4 mRNA during seed germination parallels endosperm cap weakening determined by puncture force analysis. We hypothesize that LeEXP4 is involved in the regulation of seed germination by contributing to cell wall disassembly associated with endosperm cap weakening.     

Differences in seed properties and germination between native and introduced populations of Triadica sebifera

很多入侵植物的种子比原产地植物的种子萌发的更快,这可能有利于它们成功入侵。为了理解生物入侵过程,对入侵植物的入侵种群 和原产地种群的种子特性和萌发特征同时进行评估极为必要。本研究中,我们以乌桕(Triadica sebifera)为研究对象探究种子萌发和种子 特征的种内差异。 我们利用采于2017年的乌桕种子,来自中国的12个原产地种群和美国的12个引入地种群,测定种子的物理性状(体积、重量、种皮硬度和种皮厚度)和化学特性[种仁的粗脂肪、可溶性蛋白质、糖、赤霉素(GA)和脱落酸(ABA)],并在中国的温室中测定种子的发芽率和萌发时间。此外,我们在美国用采自2016年和2017年的种子开展了研究地点(中国和美国)和种子采集年份(2016年和2017年)对种子萌发影响的比较实验。 来自引入地的种子要比来自原产地的种子萌发更快。种子的物理和化学特性测定结果表明：与原产地种子相比,引入地种子更大、种仁内含有的赤霉素浓度以及GA:ABA比更高,但粗脂肪含量较低。种子重量、种皮硬度、种皮厚度、种仁脱落酸、可溶性蛋白质和糖含量在引入地和原产地之间没有显著差异。在中国和美国温室间的种子萌发率具有相关性,但种群的萌发率具有年际变化。我们的研究结果表明,体积较大和赤霉素含量较高的种子可能有助于快速萌发,这可能也有利于乌桕在引入地的入侵。     

油菜素内酯浸种对盐胁迫番茄种子萌发的影响及其生理机制

为揭示油菜素甾醇类化合物提高作物耐盐的效应和机理,研究了10^-11、10^-10、10^-9、10^-8、10^-7、10^-6、10^-5 mol/L 2,4-表油菜素内酯（EBL）浸种处理对0、50、100、150、175 mmol/L NaCl胁迫7 d的番茄种子萌发、生长、溶质积累、抗氧化代谢的影响。结果显示：NaCl浓度越高的盐胁迫下,10^-9 mol/L EBL浸种可体现出越显著的促进番茄种子萌发的效应;在所有处理下,EBL浸种浓度过高,即10^-6、10^-5 mol/L EBL,均表现出对种子萌发的抑制效应。盐胁迫下种子萌发后,一定浓度的EBL浸种可表现出明显的增加种子胚根和下胚轴长,提高萌发种子鲜重和种子活力指数,其中10^-9 mol/L EBL浸种处理促进效果最适;EBL浸种浓度过高,则表现出抑制效应。150 mmol/L NaCl胁迫或非盐胁迫下,10^-9 mol/L EBL浸种均可降低萌发种子体内的O₂^·-、H₂O₂、丙二醛（MDA）和脯氨酸（Pro）含量;盐胁迫下,10^-9 mol/L EBL浸种可显著提高萌发种子可溶性糖（SS）和可溶性蛋白（SP）的含量。150 mmol/L NaCl胁迫或非盐胁迫下,10^-9 mol/L EBL处理可不同程度促进番茄种苗超氧化物歧化酶（SOD）和过氧化物酶（POD）活性的上升。综上所述,盐胁迫下,一定浓度范围内的EBL浸种可明显促进番茄种子萌发或成苗,其中以10^-9 mol/L EBL浸种的效果最好,主要是因为EBL施用可积极促进番茄种子萌发中物质转化,SS和SP等溶质积累增多,增强其渗透调节能力;同时SOD和POD酶活增强,缓解盐胁迫导致番茄种子萌发中的次生氧化胁迫。     

The relationship between antioxidant enzymes activity and mungbean sprouts growth during the germination of mungbean seeds treated by electrolyzed water

Slightly acidic electrolyzed water (SAEW) could promote the germination and growth of mungbean sprouts, and electron spin resonance analyses of SAEW showed that besides available chlorine, reactive oxygen species (ROS) existed in SAEW. Further research showed that, a series of antioxidant enzymes activity were also affected by SAEW. Lower superoxide dismutase activity and higher peroxidase, catalase and ascorbate peroxidase activity led to less hydrogen peroxide accumulated in mungbean sprouts treated by SAEW. Thus, the available chlorine and ROS present in the SAEW played a role in the mungbean sprouts’ antioxidant mechanism, which in turn appeared as a growth-promoting effect on the mungbean sprouts.     

皂荚天然群体间种实表型特性及种子萌发的差异分析

对位于湖北京山、湖南城步、贵州兴义、重庆秀山、四川成都、广西桂林、山东费县和甘肃天水的8个皂荚（Gleditsia sinensis Lam．）天然群体间果实和种子表型特性及种子萌发率的差异进行了研究,在此基础上,对种子特性与萌发率的相关性进行了分析。结果表明：不同皂荚群体间荚果的长度、宽度、厚度和质量以及种子长度、宽度、厚度和百粒质量均存在极显著差异（P〈O．01）;在低纬度、高降雨量和干扰程度相对大的区域产生的荚果及种子均较大。在不同温度（昼／夜温度35℃／20℃、30℃／15℃、25℃／10oC和15℃／5℃）条件下,8个皂荚群体间种子萌发率无显著差异,平均萌发率为7．70％;种皮经人为损伤后种子萌发率总体上有所提高且在不同温度条件下差异显著,其中在昼／夜温度15℃／5℃的条件下种子萌发率低于其他处理温度。在不同温度条件下,种子的长度、宽度、厚度和百粒质量与萌发率均呈正相关,但总体上相关性不显著。研究结果说明：不同皂荚群体的荚果和种子特性变异较大,且在降雨量高的地区其果实和种子较大;低温对皂荚种子的萌发有一定抑制作用;种皮损伤处理可解除皂荚种子的物理性休眠,但皂荚种子还可能存在生理休眠现象;一定程度的种子复合休眠可能是皂荚群体适应不同生境的重要生存策略之一。     

Vacuolar H(+)-ATPase is expressed in response to gibberellin during tomato seed germination

Completion of germination (radicle emergence) by gibberellin (GA)-deficient (gib-1) mutant tomato (Lycopersicon esculentum Mill.) seeds is dependent upon exogenous GA, because weakening of the endosperm tissue enclosing the radicle tip requires GA. To investigate genes that may be involved in endosperm weakening or embryo growth, differential cDNA display was used to identify mRNAs differentially expressed in gib-1 seeds imbibed in the presence or absence of GA(4+7). Among these was a GA-responsive mRNA encoding the 16-kD hydrophobic subunit c of the V(0) membrane sector of vacuolar H(+)-translocating ATPases (V-ATPase), which we termed LVA-P1. LVA-P1 mRNA expression in gib-1 seeds was dependent on GA and was particularly abundant in the micropylar region prior to radicle emergence. Both GA dependence and tissue localization of LVA-P1 mRNA expression were confirmed directly in individual gib-1 seeds using tissue printing. LVA-P1 mRNA was also expressed in wild-type seeds during development and germination, independent of exogenous GA. Specific antisera detected protein subunits A and B of the cytoplasmic V(1) sector of the V-ATPase holoenzyme complex in gib-1 seeds only in the presence of GA, and expression was localized to the micropylar region. The results suggest that V-ATPase plays a role in GA-regulated germination of tomato seeds.     

Enzyme activities during imbibition and germination of seeds of tamarack (Larix laricina)

Activities of six enzymes from extracts of separated embryos and gametophytes of tamarack [ Larix laricina (Du Roi) K. Koch] seeds were assayed at various stages of imbibition and germination. On a per seed part basis, activities of 6-phosphogluconate dehydrogenase (6-PGD, EC 1.1.1.44), glucose-6-phosphate dehydrogenase (G-6-PD, EC 1.1.1.49), malate dehydrogenase (NAD⁺–MDH, EC 1.1.1.37), isocitrate dehydrogenase (NADP⁺–IDH, EC 1.1.1.42), soluble peroxidase (PER, EC 1.11.1.7), and acid phosphatase (ACP, EC 3.1.3.2) from both the embryo and gametophyte tissues generally increased slowly, following cold stratification for 30 days and imbibition under germinating conditions for 5 days, but then increased at a faster rate with emergence of the radicle and subsequent growth of the seedling. The rate of increase of enzyme activity was highest for PER. Soluble protein levels also increased with imbibition and germination, with about 3 times greater levels present in the gametophyte than in the embryo. Heat inactivation experiments showed that, except for G-6-PD, activities were stable up to 40°C. Inactivation occurred at lower temperatures for G-6-PD, while higher temperatures were required for PER. Incubation of extracts for 7 days at 4°C indicated that loss of enzyme activity was greatest for G-6-PD (3.9% remaining) and least for PER and ACP (94 and 95% remaining, respectively).