Mechanisms associated with tolerance to flooding during germination and early seedling growth in rice (Oryza sativa)

Background and Aims

Flooding slows seed germination, imposes fatalities and delays seedling establishment in direct-seeded rice. This study describes responses of contrasting rice genotypes subjected to flooding or low oxygen stress during germination and discusses the basis of tolerance shown by certain cultivars.

Methods

In one set of experiments, dry seeds were sown in soil and either watered normally or flooded with 10 cm of water. Seedling survival and shoot and root growth were assessed and seed portions of germinating seedlings were assayed for soluble sugars and starch concentrations. The whole germinating seedlings were assayed for amylase and peroxidase activities and for ethylene production. Activities of enzymes associated with anaerobic respiration were examined and gene expression was analysed separately with seeds germinating under different amounts of dissolved oxygen in dilute agar.

Key Results

Flooding during germination reduced survival but to a lesser extent in tolerant genotypes. Starch concentration in germinating seeds decreased while sugar concentration increased under flooding, but more so in tolerant genotypes. Amylase activity correlated positively with elongation (r = 0·85 for shoot and 0·83 for root length) and with plant survival (r = 0·92). Tolerant genotypes had higher amylase activity and higher RAmy3D gene expression. Ethylene was not detected in seeds within 2 d after sowing, but increased thereafter, with a greater increase in tolerant genotypes starting 3 d after sowing. Peroxidase activity was higher in germinating seeds of sensitive genotypes and correlated negatively with survival.

Conclusions

Under low oxygen stress, tolerant genotypes germinate, grow faster and more seedlings survive. They maintain their ability to use stored starch reserves through higher amylase activity and anaerobic respiration, have higher rates of ethylene production and lower peroxidase activity as germinating seeds and as seedlings. Relevance of these traits to tolerance of flooding during germination and early growth is discussed.Key words: Amylase, anoxia, crop establishment, direct-seeded rice, ethylene, flooding, germination, hypoxia, Oryza sativa     

土壤有效硅对大豆生长发育和生理功能的影响

人工调节土壤有效硅含量及盆栽试验,研究土壤有效硅对大豆生长发育和生理功能的影响。结果表明,土壤有效硅含量在55．1～202.8mg·kg^-1范围内,随着土壤有效硅含量的提高,大豆种子萌发过程中蛋白酶和脂肪酶活性升高,淀粉酶活性无显著变化,呼吸速率加快,种子活力升高,萌发速度加快,种子萌发率无显著变化;幼苗生长过程中叶片叶绿素含量无显著变化,光合速率加快,根系活力、硝酸还原酶活力升高,蒸腾强度减弱,水分利用效率和叶含水量升高,抗旱保水能力提高。大豆幼苗含硅量与土壤有效硅含量呈线性正相关趋势(r=0．994)。土壤有效硅含量大于2028mg·kg^-1,生理功能不再显著变化,说明土壤中的硅被大豆吸收后,改善了大豆萌发种子和幼苗的生理功能,使种子萌发和幼苗生长加快。     

臭氧水浸种对黄瓜种子萌发及幼苗生长的影响

臭氧水处理10、30和50 min后的黄瓜种子,种子萌发及幼苗生长均受到促进,种子的发芽率和活力指数、萌发种子的过氧化氢酶(CAT)和淀粉酶的活性以及呼吸速率、幼苗的壮苗指数和根系活力均有提高;处理70和90min的黄瓜种子萌发和生长则受到抑制,种子电解质外渗,过氧化氢酶和淀粉酶活性下降.     

秧宝灵防止早稻烂秧的效应

从化学防寒角度,探讨防止早稻烂秧、培育壮秧的新途径。结果表明,用秧宝灵浸渍早稻种子,不仅有利于增强发芽势,提高发芽率,使种子催成根短芽壮的好秧苗,而且能有效地防止烂秧,达到培育壮秧之目的,并获得早熟增产的良好效果。     

磁化水浸种对苦瓜种子萌发及对其幼苗生理指标的影响

目的 :研究磁化水对苦瓜种子萌发的影响。方法 :用不同强度的磁化水浸种处理苦瓜种子 ,测定其以下几个生理指标 :种子的发芽率、发芽指数、活力指数和幼苗中的酯酶以及抗坏血酸的含量。实验结果显示 :磁化水能够提高苦瓜种子的发芽率、发芽指数、活力指数 ,同时也能提高苦瓜幼苗的酯酶活性 ,但对抗坏血酸含量影响不大。结论 :一定强度的磁化水能提高苦瓜种子的萌发能力 ,并使幼苗生长整齐一致。     

Respiratory Properties of Mitochondria from Rice Seedlings Germinated under Water and Their Changes during Air Adaptation

Respiratory activities were compared among rice seedlings germinated in air for 6 days (aerobic seedlings), those germinated under water for 5 days (submerged seedlings), and those grown in air for 1 day after 5 days' submerged germination (air-adapted seedlings). The respiratory activity of the submerged seedlings increased rapidly on transfer to air and reached a plateau at 16 hours in air. Respiration of the submerged seedlings was as sensitive to cyanide as those of aerobic and air-adapted seedlings. 2,4-Dinitrophenol had no effect on the respiration of the submerged seedlings, but stimulated those of the other two types of seedlings. Mitochondria from three types of seedlings did not differ in the ADP/O ratio and the respiratory control ratio (RCR) when succinate was oxidized. However, mitochondria from submerged seedlings (submerged mitochondria) showed poor RCR of about unity when malate was oxidized. Both the rate of succinate oxidation and succinate dehydrogenase activity were low in submerged mitochondria, but increased during air adaptation. Although submerged mitochondria oxidized malate very slowly, this activity increased after exposure to air without any increase in malate dehydrogenase activity. When NAD⁺ was added to submerged mitochondria, oxidation of malate was restored to the level of the aerobic controls. Addition of NAD⁺ enhanced the state 3 rate in submerged mitochondria, and RCR recovered to nearly the same value as that of the aerobic controls. Similar effects of NAD⁺ on 2-oxoglutarate oxidation were observed. All these defects in submerged mitochondria were repaired during air adaptation. These results suggest that NAD⁺-linked substrate oxidation was low in submerged mitochondria because of NAD⁺ deficiency, and that the oxidation increased with an increasing level of NAD⁺ during air adaptation.     

高吸水性种衣剂对水稻旱育秧苗生长的影响

高吸水性树脂具有较强吸水保湿功能,使用高吸水性水稻种衣剂培育旱育秧苗,可以有效控制苗床水分和湿度,同时能缓释农药和肥料,增强秧苗抗逆性。试验表明,高吸水性种衣剂处理对水稻发芽率和发芽势无不良影响,且壮根、壮苗效果显著;能减少病害发生,保持旱育秧苗发根优势和分蘖优势,达到省工省时的目的。     

壳聚糖-PVA水稻种衣剂在育秧上的应用研究

本文研究壳聚糖-PVA复合型水稻种衣剂对种子发芽率、成苗率、秧苗素质的影响以及对苗期病虫害的防治效果。结果表明,种衣剂对种子的发芽无不良影响,可提高秧苗素质,有效防止苗期病虫害发生。     

Effect of germination temperature on characteristics of phytase production from barley

The effects of germination temperature on the growth of barley seedlings for phytase production were studied at 15, 20 and 25 degrees C for 6-10 days. The growth rate of the barley seedlings was increased as the germination temperature was increased. The initial rate of total protein production was closely coupled to that of the barley growth, and the rate of total protein production tended to increase as the germination temperature was increased. SDS-PAGE analysis of total protein from the barley seedlings showed time-dependent appearance and disappearance of protein bands. Although no significant phytase activity was detected at zero time of germination, a significant increase in phytase activity up to 7.9-fold occurred during the first several days of germination then decreased. Phosphate production (viz. phytate degradation) in the barley seedlings occurred rapidly at the beginning of germination. However, the rate of production continued to decrease with further germination. A time lag of about 1-2 days between the rate of total protein production and that of phytase production was observed. Unlike the extent of total protein production, that of phytase production was similar irrespective of germination temperature. Partial purification of a crude enzyme extract by hydrophobic interaction chromatography resulted in two phytase fractions (PI and PII). Zymogram analysis demonstrated that PI had two bands with molecular masses of about 66 and 123 kDa while PII had one band corresponding to a molecular mass of about 96 kDa. The optimal temperature for PI was found to be 55 degrees C, while it was 50 degrees C for PII. The enzyme fraction PI had a pH optimum at 6.0, whereas the optimum pH for PII was found to be 5.0. Addition of 0.1% (v/v) Tween 80 was found to increase enzyme activity significantly (i.e., 167% for PI and 137% for PII). Phytate in cereals including barley, rice, corn and soybean degraded effectively by the treatment of the barley phytases.     

Effect of Barbiturates on Rice Seedling Germination and Developmental Behaviour

Rice seedling germination and developmental behaviour were examinedin the presence of six different barbituric acid derivatives.The relationship between liquid/aqueous partition coefficientsof barbiturates and their interference with, but not completeinhibition of, normal mitochondria1 oxidative metabolism observedin animal mitochondria in vitro also appear to be factors influencingthe effects of these compounds on the germination and developmentalbehaviour of the rice seedlings described in this investigation.Evidence is presented here which points to normal oxidativemetabolism as being a necessary requisite for normal light responseand normal morphogenetic development in germinating rice seedlings.     

成熟度与烘干温度对结球甘蓝种子质量的影响

以结球甘蓝品种冬升种子为材料,研究了不同成熟度和烘干温度下种子秕粒率、千粒重、发芽率、生理活性情况以及不同烘干温度下种子的含水量.结果表明,结球甘蓝冬升开花后45～55 d采收的种子,发芽率均达到了95%以上;随着种子成熟度提高,其种子质量、发芽活力及其超氧化物歧化酶(SOD)、过氧化物酶(POD)、脱氢酶活性显著上升,而相对电导率显著下降.与对照(自然风干)相比,30～50℃的烘干温度对种子千粒重和秕粒率无显著性影响,也仅在50℃下可使种子的发芽活力显著降低;随着烘干温度的升高,种子的SOD、POD和脱氢酶活性逐渐显著下降,相对电导率则逐渐显著上升;30～50℃烘干6 h种子的含水量由13.3%降至5.4%左右.研究发现,结球甘蓝冬升开花授粉后45 d种子已达到了采收程度,30～50℃烘干6 h种子含水量已达到储藏要求,并且愈接近自然干燥温度(30～40℃)的处理,种子发芽能力愈好;甘蓝种子活力与其SOD、POD和脱氢酶活性呈正相关,而与其相对电导率呈负相关.     

Drought resistance in rice seedlings conferred by seed priming

Seed priming is a method by which seeds are subjected to different stress conditions to impart stress adaptation in seedlings germinating and growing under stressful situations. Drought stress is a major reason behind failure of crops. We studied the effects of hydropriming, dehydration priming (induced by PEG), and osmopriming (induced by NaCl and KH₂PO₄) on subsequent germination, growth and anti-oxidant defense mechanisms of 2-week-old rice seedlings under continuing dehydration stress. Unprimed seeds grown in PEG showed significantly lower germination and growth along with significantly higher reactive oxygen species (ROS) and lipid peroxidation levels. Among the priming methods, 5 % PEG priming was found to be the best in terms of germination and growth rate along with the lowest amount of ROS and lipid peroxidation (malondialdehyde [MDA]) values. MDA levels were reduced significantly by all of the priming methods. Hence, reduction of lipid peroxidation may be a key factor underlying the drought tolerance produced by the priming treatments. Glutathione peroxidase (GPX) activity seemed to bear an excellent correlation with oxidative stress resistance through seed priming. The PEG priming produced minimum peroxidative damage and superior germination and growth rate along with efficient GPX activity, overexpressed MnSOD and maintenance of HSP70 expression in normal as well as in drought condition. Therefore, in PEG-primed seeds the existence of robust protective mechanisms is definitely indicated.     

Synthesis and biological activity of amino acid conjugates of abscisic acid

We prepared 19 amino acid conjugates of the plant hormone abscisic acid (ABA) and investigated their biological activity, enzymatic hydrolysis by a recombinant Arabidopsis amidohydrolases GST-ILR1 and GST-IAR3, and metabolic fate in rice seedlings. Different sets of ABA-amino acids induced ABA-like responses in different plants. Some ABA-amino acids, including some that were active in bioassays, were hydrolyzed by recombinant Arabidopsis GST-IAR3, although GST-ILR1 did not show hydrolysis activity for any of the ABA-amino acids. ABA-L-Ala, which was active in all the bioassays, an Arabidopsis seed germination, spinach seed germination, and rice seedling elongation assays, except in a lettuce seed germination assay and was hydrolyzed by GST-IAR3, was hydrolyzed to free ABA in rice seedlings. These findings suggest that some plant amidohydrolases hydrolyze some ABA-amino acid conjugates. Because our study indicates the possibility that different plants have hydrolyzing activity toward different ABA-amino acids, an ABA-amino acid may function as a species-selective pro-hormone of ABA.     

The Germination of Carrot (Daucus carota L.) Seed Harvested on Two Dates: A Physiological and Biochemical Study

The germination of two batches of carrot seed, harvested 44and 104 d after anthesis, was compared at 10 °C. Proteinand nucleic acid contents of the seeds were measured at intervals,together with fresh weight and respiration rate. The matureseed germinated 3.7 d earlier than the immature seed, with nodifference in percentage germination. The dry mature seed containedmore protein and nucleic acid per unit dry matter than did theimmature seed, and proportions of nucleic acid present as rRNAand poly(A)RNA were greater in the mature seed. The sequenceof metabolic reactivation was the same in both batches of seed,as were the relative rates of increase of each nucleic acidcomponent, fresh weight, and respiration rate. Differences inthe composition of the dry seed appear to be responsible forthe observed difference in rate of germination between the twoseed batches.     

多裂骆驼蓬提取物对黄瓜种子萌发和幼苗生理特性的影响

通过室内培养和盆栽土培试验研究了多裂骆驼蓬提取物对黄瓜种子萌发和幼苗生长及生理特性的影响。结果表明,多裂骆驼蓬总生物碱提取液、水溶性生物碱提取液和脂溶性生物碱提取液浸种均抑制黄瓜种子萌发过程中淀粉酶、蛋白酶和脂肪酶活性,种子活力和萌发速率降低,呼吸速率减慢;幼苗生长过程中根系活力、硝酸还原酶活性升高,叶绿素含量增加,超氧化物歧化酶（SOD）和过氧化物酶（POD）活性提高。说明用多裂骆驼蓬提取液浸种能够促进黄瓜幼苗生长,有利于培育壮苗。     

Bleach-induced Germination and Breakage of Dormancy of Seeds of Alectra vogelii

The effects of solutions of sodium hypochlorite and calcium hypochlorite (bleach) on dormancy and germination of Alectra vogelii seeds were investigated. Dry (non-pretreated) and 10-day water-pretreated seeds were exposed to various bleach concentrations, as well as to the host (Vigna unguiculata) root exudate. The 15-month-old Alectra seeds used were partially dormant in that only 40% or less of the 10-day pretreated seeds could be stimulated to germinate by the standard host root exudate. Comparable percentage germination of nonpretreated seeds was achieved with certain concentrations of bleach and the halogens, chlorine and bromine. Bleach and the halogens served not only as germination stimulant for pretreated Alectra seeds but appropriate concentrations induced also high percentage (70–90%) germination indicating a breakage of seed dormancy as well. The activity of the bleach in stimulating high percentage germination could be significantly reduced when the optimally bleach-treated seeds were rinsed daily with water during the germination period. Vigna root exudate, but not bleach, was shown to act as root stimulant for Alectra seedlings. The possible mechanism of the bleach effect on germination and dormancy of Alectra seeds is discussed.     

Photosynthetic Development of Anaerobically Grown Rice (Oryza sativa) after Exposure to Air

During anaerobic germination, rice produces a coleoptile devoid of carotenoid and chlorophyll. Further development and greening of the shoot occur upon exposure of the seedlings to air. In this study, a comparison was made between anaerobically (N₂) germinated rice, greened upon exposure to air, and air/dark (A/D) germinated seedlings, greened upon exposure to light. After exposure to air, N₂-grown seedlings had a 76-hour lag before net oxygen evolution occurred compared to a 6-hour lag for A/D-grown seedlings. After 98 h of greening, N₂-grown seedlings reached a rate of oxygen evolution equivalent to that of A/D-grown seedlings after 24 hours. Chlorophyll and carotenoid content showed a similar lag, but did not reach the level found in A/D-grown seedlings even after 124 hours of exposure to air. RuBPcase activity also lagged in N₂-grown seedlings, ultimately reaching greater values than in the `greened' A/D-grown seedlings. Phosphoenolpyruvate carboxylase activity was constant and low in all treatments except for a transient increase after 24 hours of greening of the N₂-grown seedlings.     

Effect of Radiation Quality on Growth and Photosynthesis of Acacia mangium Seedlings

Radiation quality was an important environmental cue to stimulate seed germination in Acacia mangium. The photo-synthetic CO₂ assimilation rate, dark respiration rate, total biomass, and relative growth rate of seedlings grown under monochromatic radiation were significantly lower than those of seedlings grown under full spectrum radiation. Blue and red radiation induced shade-avoidance and shade-tolerant responses of A. mangium seedlings, respectively.     

Allelopathy of rice (Oryza sativa L.) root exudates and its relations with Orobanche cumana Wallr. and Orobanche minor Sm. germination

Allelopathy has been considered not only as an environmentally friendly approach for weed control but also a potential reason causing autotoxicity in crop production. In this study, the responses of seeds of lettuce, wheat, rice, clover broomrape (CB), and sunflower broomrape (SB) to the root exudates of rice cultivars were studied. Lettuce germination was promoted by root exudates of Yliangyou 3218 and I-Kung-Pao. Wheat seedling growth was inhibited by all nine rice species. I-Kung-Pao and Ganxin 203 exerted greater autotoxicity than other cultivars. Yongyou 15 and I-Kung-Pao induced the highest germination rate of CB, while Yongyou 13, Zhongzao 22, and I-Kung-Pao induced the highest germination rate of SB. A significantly correlation was noted between germination-inducing ability on broomrape seeds and allelopathic effects on target plants. It is suggested that using broomrape seeds germination is a better receptor for the identification of rice allelopathic potential.     

Inhibitory Effects of Cycasin on Germination,Growth and α-Amylase Biosynthesis in Rice Plants

Cycasin, the toxic glycoside of cycad plants, interfered with seed germination and seedling growth of Gramineae, Crucifereae and Leguminosae. The shoots and roots of seedlings showed wilting, chlorosis and necrosis. Rice plants were most sensitive and soybean plants rather tolerant.

Respiration and α-amylase activity were markedly low in the rice seedlings treated with cycasin. Both cycasin and its aglycone, methylazoxymethanol, did not inhibit the activity of α-amylase, but did suppress the formation of α-amylase in rice endosperms. Exogenous gibberellin considerably reversed the inhibition of germination and growth, and the suppression of α-amylase formation caused by these toxins.