Caffeic acid affects early growth, and morphogenetic response of hypocotyl cuttings of mung bean (Phaseolus aureus)

Caffeic acid (CA) is one of the most common cinnamic acids ubiquitously present in plants and implicated in a variety of interactions including allelopathy among plants and microbes. This study investigated the possible interference of CA with root growth and the process of rhizogenesis in hypocotyl cuttings of mung bean (Phaseolus aureus=Vigna radiata). Results indicated that CA (0-1000 microM) significantly suppressed root growth of mung bean, and impaired adventitious root formation and root length in the mung bean hypocotyl cuttings. Further investigations into the role of CA in hampering root formation indicated its interference with the biochemical processes involved in rooting process at the three stages - root initiation (third day; RI), root expression (fifth day; RE), and post-expression (seventh day; PE) - of rhizogenesis. CA caused significant changes in the activities of proteases, peroxidases (PODs), and polyphenol oxidases (PPOs) during root development and decreased the content of total endogenous phenolics (TP) in the hypocotyl cuttings. The enhanced activity of PODs and PPOs, though, relates to lignification and/or phenolic metabolism during rhizogenesis; yet their protective role to CA-induced stress, especially during the PE phase, is not ruled out. At 1000 microM CA, where rooting was significantly affected, TP content was very high during the RI phase, thus indicating its non-utilization. The study concludes that CA interferes with the rooting potential of mung bean hypocotyl cuttings by altering the activities of PODs and PPOs and the endogenous TP content that play a key role in rhizogenesis.     

2-Benzoxazolinone (BOA) induced oxidative stress, lipid peroxidation and changes in some antioxidant enzyme activities in mung bean (Phaseolus aureus)

2-Benzoxazolinone (BOA), a well-known allelochemical with strong phytotoxicity, is a potential herbicidal candidate. The aim of the present study was to determine whether phytotoxicity of BOA is due to induction of oxidative stress caused by generation of reactive oxygen species (ROS) and the changes in levels of antioxidant enzymes induced in response to BOA. Effect of BOA was studied on electrolyte leakage, lipid peroxidation (LP), hydrogen peroxide (H(2)O(2)) generation, proline (PRO) accumulation, and activities of antioxidant enzymes-superoxide dismutase (SOD, 1.15.1.1), ascorbate peroxidase (APX, 1.11.1.11), guaiacol peroxidase (GPX, 1.11.1.7), catalase (CAT, 1.11.1.6) and glutathione reductase (GR, 1.6.4.2) in Phaseolus aureus (mung bean). BOA significantly enhanced malondialdehyde (MDA) content, a product of LP, in both leaves and roots of mung bean. The amount of H(2)O(2), a product of oxidative stress, and endogenous PRO increased many-fold in response to BOA. Accumulation of PRO, MDA and H(2)O(2) indicates the cellular damage in the target tissue caused by ROS generated by BOA. In response to BOA, there was a significant increase in the activities of scavenging enzymes SOD, APX, GPX, CAT, and GR in root and leaf tissue of mung bean. At 5 mM BOA, GR activity in roots showed a nearly 22-fold increase over that in control. The present study concludes that BOA induces oxidative stress in mung bean through generation of ROS and upregulation of activities of various scavenging enzymes.     

Effects of 2-benzoxazolinone on the germination, early growth and morphogenetic response of mung bean (Phaseolus aureus)

2‐Benzoxazolinone (BOA), a type of hydroxamic acid present in cereals and implicated in allelopathy, is now being viewed as a potential candidate for the development of natural herbicides. A study was conducted to determine the effect of BOA on mung bean (Phaseolus aureus) through a multitude of bioassays to understand its physiological and biochemical action. It was observed that BOA significantly decreased the germination of mung bean and its early growth (measured in terms of seedling length and dry weight). A typical dose–response relationship was observed with BOA treatment, and I₅₀ values (concentrations at which 50% inhibition occurs) for germination, seedling length and seedling dry weight were calculated to be 4.3, 0.71 and 0.77 mM , respectively. There was therefore a greater inhibitory effect on seedling growth than on germination. Treated seedlings were characterised by a loss of chlorophyll and decreased respiratory activity, indicating a possible adverse effect of BOA on photosynthetic and respiratory metabolism. Mitotic activity in root‐tip cells of onion (Allium cepa) was completely arrested in response to BOA treatment, and the cells exhibited abnormality in shape and size. BOA also adversely affected rhizogenesis in hypocotyl cuttings of mung bean, indicating an impact on morphogenetic potential. It was associated with significant changes in the protein content and activities of proteases and polyphenol oxidases during the root development phase. This study concludes that BOA interfered with essential biochemical processes in mung bean. Such studies provide useful information on the biochemical and physiological modes of actions of BOA, with a view to its use as a herbicidal compound.     

Ferulic acid impairs rhizogenesis and root growth,and alters associated biochemical changes in mung bean (Vigna radiata) hypocotyls

The present study investigated the effect of ferulic acid (FA; 0–1000 µM) on early growth, and rhizogenesis in mung bean (Vigna radiata) hypocotyls and associated biochemical changes. FA severely affected the radicle elongation and number of secondary roots after 72 h. The root and shoot length, number and length of secondary roots, and seedling dry weight of one-week-old seedlings of mung bean were decreased by 64%. The rooting potential (percent rooting, number and length of adventitious roots) of mung bean hypocotyls under in vitro conditions was significantly inhibited in response to 1–100 µM FA. At 1000 µM there was complete cessation of rooting. FA caused a reduction in the contents of water-soluble proteins and endogenous total phenolics, whereas the activities of proteases, peroxidases, and polyphenol peroxidases increased. The study concludes that FA inhibits root growth and development, and in vitro rooting process in mung bean by interfering with biochemical processes that are crucial for root formation.     

Lead (Pb)-Inhibited Radicle Emergence in Brassica campestris Involves Alterations in Starch-Metabolizing Enzymes

Lead (Pb) is a toxic heavy metal released into the natural environment and known to cause oxidative damage and alter antioxidant mechanism in plants. However, not much is known about the interference of Pb with the biochemical processes and carbohydrate metabolism during seed germination. We, therefore, investigated the effect of Pb (50-500 μM) upon biochemical alterations in germinating seeds (at 24-h stage) of Brassica campestris L. Pb treatment significantly enhanced protein and carbohydrate contents that increased by ~43% and 200%, respectively, at 500-μM Pb over control. In contrast, the activities of starch/carbohydrate-metabolizing enzymes--α-amylases, β-amylases, acid invertases, and acid phosphatases--decreased by ~54%, 60%, 74%, and 52%, respectively, over control. Activities of peroxidases and polyphenol oxidases, involved in stress acclimation, however, increased by ~1.2- to 3.9-folds and 0.4- to 1.4-folds upon 50-500-μM Pb treatment. Pb enhanced oxidizing ability by 10 to 16.7 times over control suggesting interference with emerging root's oxidizing capacity. The study concludes that Pb exposure inhibits radicle emergence from B. campestris by interfering with the biochemical processes linked to protein and starch metabolism.     

Phytotoxic effects of β-pinene on early growth and associated biochemical changes in rice

β-Pinene, an oxygenated monoterpene, is one of the major monoterpenes emitted into the atmosphere from forest areas and trees. Besides, it is a principal component of essential oils of a number of aromatic plants, which are involved in a variety of ecological interactions, including allelopathy, in the natural environment. However, studies pertaining to phytotoxicity and biochemical effect(s) of β-pinene are largely lacking. We investigated the effect of β-pinene (0.02, 0.04, 0.08, 0.20, 0.40 and 0.80 mg/ml) in a dose- and time-dependent manner on early seedling growth, dry weight accumulation, photosynthetic pigments and changes in macromolecule (protein and carbohydrate) content and activities of enzymes—proteases, α- and β-amylases, polyphenol oxidases and peroxidases- in rice (Oryza sativa) after 3rd, 5th and 7th day of exposure. β-pinene (≥0.04 mg/ml) significantly reduced the root (by 13–87%) and coleoptile (by 5–80%) length of rice. Exposure to β-pinene reduced total chlorophyll content in rice coleoptiles suggesting a negative impact on photosynthesis. The content of macromolecules (proteins and carbohydrates) enhanced significantly in response to β-pinene, whereas the activities of hydrolyzing enzymes—proteases, α-amylases, and β-amylases—declined (by 30–85, 26–84, 27–74%, respectively) in β-pinene-exposed seedlings. In contrast, the activities of peroxidases (POX) and polyphenol oxidases (PPO) enhanced significantly (by 16–152 and 53–290%, respectively) in rice roots in response to β-pinene in a dose- and time-dependent manner. Increased activities of POX and PPO indicate their involvement in providing protection and/or conferring resistance against β-pinene-induced stress. The study concludes that β-pinene inhibits the early growth of rice by altering the plant biochemical status and enhancing activities of POXs and PPOs involved in general plant defense.     

Effect of Paraquat on Tomato Roots Cultured in Vitro Susceptible and Resistant to Nematodes

Roots from two tomato cultivars, one resistant (Rossol) andthe other susceptible (Roma VF) to root-knot nematodes, werecultured in vitro and tested for their response to paraquat,an herbicide which generates superoxide in cells. The effectof paraquat on the permeability of root membranes was testedby the safranine method. Membranes from resistant roots wereinjured more markedly than susceptible ones. A statistical analysisof the variations on a series of enzyme activities from rootsincubated for 2 days with paraquat was carried out. Superoxidedismutase was repressed in treated resistant roots togetherwith ascorbate and glutathione peroxidases. In susceptible rootsthese enzymes were not significantly affected by paraquat treatmentexcept for ascorbate peroxidase which increased slightly. Syringaldazineand polyphenol oxidases, two enzymes active in the hypersensitiveresponse to nematodes, increased in resistant but not in thesusceptible cultivar. p-Phenilendiamine-pyrocathecol oxidasewas not affected while catalase increased markedly by paraquattreatment in both cultivars. (Received January 21, 1991; Accepted August 6, 1991)     

莲根在个体发育中表现出的适应

本文研究了莲根和根第在个体发育中的发生与发育及其适应。苗端发育先于根端;节生根首先从胚芽第一节发生。很少的种子在种子萌发时有胚根原基的微弱发育,但从未见胚根突出种皮。须根系仅由节生根组成而根端终生不发育成直根系,该须根系应称为原生须根系。其特点是在一个生长季节之内死亡,是一种新的发育类型,特命为年内全更新型。     

Influence of enzymes on the quality of processed vegetables and fruits

These organic catalysts—peroxidases, polyphenol oxidases and pectic enzymes among them—must be inactivated by heat or chemical treatment if high-quality food products are to be stored. Knowledge of their activity and control is still meagre.     

l-DOPA (l-3,4-dihydroxyphenylalanine) affects rooting potential and associated biochemical changes in hypocotyl of mung bean, and inhibits mitotic activity in onion root tips

A study was undertaken to explore the effect of l-DOPA (l-3,4-dihydroxyphenylalanine) on the rooting potential of hypocotyl cuttings of mung bean (Phaseolus aureus Roxb. var. SML-32) and related biochemical changes at the post-expression phase. At lower concentrations of (0.0001–0.1 mM) l-DOPA, there was no change in rooting potential, though the average number of roots per cutting and root length were significantly decreased (except at 0.0001 mM). However, at 1.0 mM concentration, a 50% inhibition in rooting potential was noticed and the root number and length were severely reduced. In contrast, at 2.5 mM l-DOPA, none of the hypocotyl cutting rooted. The decrease in rooting potential was associated with a significant effect on the biochemical changes measured in terms of protein and carbohydrate metabolism and activity of peroxidases. In the l-DOPA treated hypocotyl cuttings, there was a significant reduction in the protein and carbohydrate content, whereas activities of associated enzymes proteases and amylases decreased, particularly at higher treatment concentration (>1.0 mM). It indicated negative effect of l-DOPA on these two important metabolic processes. Likewise, activity of peroxidases also decreased in the l-DOPA treated hypocotyl mung bean cuttings thereby indicating its role in suppressing rhizogenesis as the enzyme is involved in lignification process during cell division. l-DOPA suppressed mitotic activity in the root tip cells of onion indicating thereby its interference with the cell division, which is required for the formation of new meristematic tissue during rhizogenesis. Based on the obtained results, it is concluded that l-DOPA interferes with the various biochemical processes in the mung bean hypocotyl cuttings thereby affecting their rooting potential.     

Embryonal growth-correlative effects in pea seedlings as reflected in the level of endogenous gibberellin-like and cytokinin-like substances

The level of endogenous gibberellin-like and cytokinin-like substances was estimated in the plumule and the root of 2-to 3-day-old intact pea seedlings, in the plumule of seedlings from which the radicle was cut off, and in the root of seedlings from which the plumule was removed. An increase in the level of gibberellin-like substances and a decrease in the level of cytokininlike substances in the plumule as a consequence of radicle amputation were observed within 48 h. Plumule amputation resulted in a decreased level of both gibberellin-like and cytokinin-like substances in the root in the same period.     

亚精胺浸种对渗透胁迫下白三叶子萌发及淀粉代谢的影响

以‘拉丁诺’白三叶为材料,用0、10％、15％、20％（W/V）即的聚乙二醇（PEG-6000）溶液模拟干旱条件,研究亚精胺fSpd）浸种对渗透胁迫下白三叶种子萌发和淀粉代谢的影响。结果表明,在PEG渗透胁迫下,白三叶种子的发芽率、发芽势、发芽指数、胚芽及胚根鲜重和胚根长度均显著（P〈0．05）降低,淀粉水解为糖类的速率减慢;与蒸馏水浸种相比,0．05mmol．L-1 Spd浸种处理显著（P〈0．05）提高了在渗透胁迫条件下种子的发芽率、发芽势、发芽指数、胚芽及胚根鲜重、干重和胚根长度,同时大幅提高了α-淀粉酶、β-淀粉酶及（α＋β）．淀粉酶总活性,降低了淀粉含量,增加了还原糖和葡萄糖含量。说明Spd浸种提高了白三叶种子在渗透胁迫下的萌发能力和幼苗生长的环境适应性,这可能与增强种子体内淀粉酶活性,加速淀粉水解为还原糖和葡萄糖,为种子萌发和幼苗早期生长及时提供充足能量有关。     

Cr(VI) Imposed Toxicity in Maize Seedlings Assessed in Terms of Disruption in Carbohydrate Metabolism

The present study examined the toxic effects of Cr(VI; 100, 250 and 500 μM) in maize seedlings by investigating the changes in carbohydrate metabolism after 48, 96, and 144 h of exposure. Cr-stress results in severe alterations in the contents of carbohydrates and reducing sugars and the activities of carbohydrate metabolizing enzymes, amylases, phosphatases and phosphorylases, and invertases in maize seedlings. Under Cr stress, the contents of carbohydrates and reducing sugars declined in roots, whereas an increase was noticed in leaves. The catalytic activity of carbohydrate metabolizing enzymes, except invertases, in roots declined in the presence of Cr(VI) in a concentration- and exposure time-dependent manner. In contrast, the activities of these enzymes were enhanced in leaves under Cr(VI) stress. The activity of invertases increased with increasing amount of Cr(VI) but declined with an increase in the time interval. In conclusion, our results show that carbohydrate metabolism is severely affected under Cr(VI) toxicity. The study suggests that Cr-induced perturbations in the carbohydrate metabolism are one of the factors resulting in growth inhibition under Cr(VI) stress.     

两株链霉菌对玉米的促生增产作用及机理

为探索娄彻氏链霉菌(D74)和密旋链霉菌(Act12)及其混合菌剂对玉米生长的影响,采用皿内发芽试验、沙培试验及小区试验观察供试链霉菌无细胞发酵滤液及活菌制剂种子包衣对玉米生物学性状、叶片诱导酶活性、光合作用、穗性状、产量及品质的影响.结果表明: 供试链霉菌无细胞发酵滤液处理玉米种子可促进玉米胚根、胚轴及幼苗生长,提高玉米幼苗叶片诱导酶活性.D74发酵液稀释1000倍处理可使玉米胚根长度、胚轴长度及须根数较对照分别增加43.4%、26.4%和100.7%(P＜0.05);D74原液可使玉米叶片多酚氧化酶(PPO)活性较对照增加40.2%.Act12发酵液稀释100倍处理可使玉米胚根长度、胚轴长度和须根数较对照分别增加36.3%、36.3%和117.5%(P＜0.05),玉米幼苗总鲜质量、根系鲜质量分别较对照增加31.1%、36.6%(P＜0.05);Act12稀释10及1000倍处理玉米种子可使玉米叶片PPO活性分别较对照增加38.1%和39.5%(P＜0.05).混合菌剂种子包衣具有以下作用:1)促进玉米根系发育;2)显著增强叶片光合能力;3)显著改善穗性状并提高籽粒产量;4)显著促进籽粒灌浆速度;5)明显提高灌浆期玉米叶片诱导酶活性.表明供试链霉菌制剂包衣玉米种子可显著影响玉米的生物学特性、光合生理及生化代谢,刺激根系发育,促进玉米生长,提高产量.     

Germination, seedling morphology and establishment of Cotnbretum bauchiense Hutch. & Dalz. (Combretaceae)

ONYEKWELU, S. S. C, 1990. Germination, seedling morphology and establishment of Cotnbretum bauchiense Hutch. & Dalz. (Combretaceae). Cotnbretum bauchiense is a suffrutex with short, erect, usually herbaceous stems arising from a woody root stock. It appears in savanna soon after fire and (lowers within a few weeks. The fruits germinate in 5–6 days. The germination is cryptogeal. On germination the true radicle and the apparent radicle formed by fused cotyledon stalks push down into the soil, carrying the plumule with them. The cotyledon lamina and part of the fused cotyledon stalks remain above the soil. Both the apparent radicle and the true radicle produce roots. Below the soil at the joint of the apparent radicle and the true radicle the plumule produces 1–3 shoots which grow out to the surface of the soil. The underground portion of the shoot bears scale leaves, from which the plant regenerates when the aerial shoot is damaged by fire. This type of germination is an adaptation that ensures successful establishment in an environment that is subjected to fire.     

Uptake and translocation of phytochemical 2-benzoxazolinone (BOA) in radish seeds and seedlings

The molecular aspects of phytochemical interactions between plants, especially the process of phytochemical translocation by the target plant, remain challenging for those studying allelopathy. 2-Benzoxazolinone (BOA) is a natural chemical produced by rye (Secale cereale) and is known to have phytotoxic effects on weed seeds and seedlings. The translocation of BOA into target plants has been poorly investigated. Therefore, the total absorption of [ring U 14C] BOA was estimated by oxidizing whole seedlings of Raphanus sativus cv. for 8 days and quantifying the radioactivity. Non-radiolabelled BOA in seedlings was also estimated by HPLC. BOA applied at 10(-3) M was readily taken up by germinated radish at a rate of 1556 nmol g(-1) FW. At these same concentrations, BOA reduced radish germination by 50% and caused a delay in radicle elongation. Exogenous BOA was responsible for the observed germination inhibition. At a concentration of 10(-5) M, BOA was taken up by germinated seeds (31 nmol g(-1) FW), but this quantity did not affect radish germination. Labelled BOA was not mineralized in the culture medium during seedling growth as no 14CO2 was recovered. Both 10(-3) and 10(-5) M BOA were translocated into radish organs, mainly into roots and cotyledons. These organs were then identified as potential physiological target sites. Cotyledons remained the target sink (44% of the total radioactivity). The kinetics of BOA uptake at 10(-3) and 10(-5) M in radish seedlings was identical: BOA accumulation was proportional to its initial concentration. A comparison between radioactivity and HPLC quantification for 10(-3) M BOA indicated that BOA (along with some metabolites) could effectively be recovered in radish organs using chromatography.     

Pre-treatment of seeds with static magnetic field ameliorates soil water stress in seedlings of maize (Zea mays L.)

The effect of magnetic field (MF) treatments of maize (Zea mays L.) var. Ganga Safed 2 seeds on the growth, leaf water status, photosynthesis and antioxidant enzyme system under soil water stress was investigated under greenhouse conditions. The seeds were exposed to static MFs of 100 and 200 mT for 2 and 1 h, respectively. The treated seeds were sown in sand beds for seven days and transplanted in pots that were maintained at -0.03, -0.2 and -0.4 MPa soil water potentials under greenhouse conditions. MF exposure of seeds significantly enhanced all growth parameters, compared to the control seedlings. The significant increase in root parameters in seedlings from magnetically-exposed seeds resulted in maintenance of better leaf water status in terms of increase in leaf water potential, turgor potential and relative water content. Photosynthesis, stomatal conductance and chlorophyll content increased in plants from treated seeds, compared to control under irrigated and mild stress condition. Leaves from plants of magnetically-treated seeds showed decreased levels of hydrogen peroxide and antioxidant defense system enzymes (peroxidases, catalase and superoxide dismutase) under moisture stress conditions, when compared with untreated controls. Mild stress of -0.2 MPa induced a stimulating effect on functional root parameters, especially in 200 mT treated seedlings which can be exploited profitably for rain fed conditions. Our results suggested that MF treatment (100 mT for 2 h and 200 for 1 h) of maize seeds enhanced the seedling growth, leaf water status, photosynthesis rate and lowered the antioxidant defense system of seedlings under soil water stress. Thus, pre sowing static magnetic field treatment of seeds can be effectively used for improving growth under water stress.     

Changes in protein profile of pigeonpea genotypes in response to NaCl and boron stress

Two pigeonpea [Cajanus cajan (L.) Millsp.] genotypes, a salt tolerant Manak and a salt sensitive ICPL 88039 were subjected to stress treatment of 3 mM boron, 60 mM NaCl and boron + NaCl at the seedling stage. Radicle and plumule proteins were analyzed by SDS-PAGE. Boron treatment increased 28.3 kDa proteins in plumule and 38.3 and 51.9 kDa proteins in radicle of Manak, however, there was no specific protein in ICPL 88039 either in plumule or in radicle. In NaCl treatment 95.6 kDa proteins appeared in plumule and 67.5 kDa proteins in radicle of Manak. Conversely content of some proteins decreased by boron treatment alone or in combination with NaCl although they were present in the controls. Thus, 54.3 kDa protein disappeared in ICPL 88039 plumule, 68.4 kDa in Manak radicle and 28.1 kDa in ICPL 88039 radicle.     

Activity of tocopherol oxidase in Phaseolus coccineus seedlings

In the present study, we have demonstrated that membrane-free extracts of etiolated shoots of Phaseolus coccineus seedlings show tocopherol oxidase activity. For this reaction, presence of membrane lipids, such as lecithin and mixture of plant lipids was required. The rate of the reaction was the highest for α-tocopherol and decreased in the order α ? β > γ > δ tocopherols. In the case of α-tocopherol, the main oxidation product was α-tocopherolquinone, while for the other tocopherol homologues the dominant products were other derivatives. When the enzyme activity was measured in leaves, hypocotyls and roots of etiolated seedlings of P. coccineus, the oxidase activity was the highest in extracts of leaves and decreased towards the roots where no activity was detected. The effect of hydrogen peroxide and of different inhibitors on the reaction suggest that tocopherol oxidase does not belong to peroxidases or flavin oxidases but rather to multi-copper oxidases, such as polyphenol oxidases or laccases. On the other hand, catechol, the well-known substrate of polyphenol oxidases and laccases, was not oxidized by the enzyme, indicating a high substrate specificity of the tocopherol oxidase.