Chuanjiu He

Transduction of an Ethylene Signal Is Required for Cell Death and Lysis in the Root Cortex of Maize during Aerenchyma Formation Induced by Hypoxia

Ethylene has been implicated in signaling cell death in the lysigenous formation of gas spaces (aerenchyma) in the cortex of adventitious roots of maize (Zea mays) subjected to hypoxia. Various antagonists that are known to modify particular steps in signal transduction in other plant systems were applied at low concentrations to normoxic and hypoxic roots of maize, and the effect on cell death (aerenchyma formation) and the increase in cellulase activity that precedes the appearance of cell degeneration were measured. Both cellulase activity and cell death were inhibited in hypoxic roots in the presence of antagonists of inositol phospholipids, Ca2+- calmodulin, and protein kinases. By contrast, there was a parallel promotion of cellulase activity and cell death in hypoxic and normoxic roots by contact with reagents that activate G-proteins, increase cytosolic Ca2+, or inhibit protein phosphatases. Most of these reagents had no effect on ethylene biosynthesis and did not arrest root extension. These results indicate that the transduction of an ethylene signal leading to an increase in intracellular Ca2+ is necessary for cell death and the resulting aerenchyma development in roots of maize subjected to hypoxia.

Ethylene Biosynthesis during Aerenchyma Formation in Roots of Maize Subjected to Mechanical Impedance and Hypoxia

Germinated maize (Zea mays L.) seedlings were enclosed in modified triaxial cells in an artificial substrate and exposed to oxygen deficiency stress (4% oxygen, hypoxia) or to mechanical resistance to elongation growth (mechanical impedance) achieved by external pressure on the artificial substrate, or to both hypoxia and impedance simultaneously. Compared with controls, seedlings that received either hypoxia or mechanical impedance exhibited increased rates of ethylene evolution, greater activities of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase, ACC oxidase, and cellulase, and more cell death and aerenchyma formation in the root cortex. Effects of hypoxia plus mechanical impedance were strongly synergistic on ethylene evolution and ACC synthase activity; cellulase activity, ACC oxidase activity, or aerenchyma formation did not exhibit this synergism. In addition, the lag between the onset of stress and increases in both ACC synthase activity and ethylene production was shortened by 2 to 3 h when mechanical impedance or impedance plus hypoxia was applied compared with hypoxia alone. The synergistic effects of hypoxia and mechanical impedance and the earlier responses to mechanical impedance than to hypoxia suggest that different mechanisms are involved in the promotive effects of these stresses on maize root ethylene biosynthesis.

Induction of Enzymes Associated with Lysigenous Aerenchyma Formation in Roots of Zea mays during Hypoxia or Nitrogen Starvation

Either hypoxia, which stimulates ethylene biosynthesis, or temporary N starvation, which depresses ethylene production, leads to formation of aerenchyma in maize (Zea mays L.) adventitious roots by extensive lysis of cortical cells. We studied the activity of enzymes closely involved in either ethylene formation (1-amino-cyclopropane-1-carboxylic acid synthase [ACC synthase]) or cell-wall dissolution (cellulase). Activity of ACC synthase was stimulated in the apical zone of intact roots by hypoxia, but not by anoxia or N starvation. However, N starvation, as well as hypoxia, did enhance cellulase activity in the apical zone, but not in the older zones of the same roots. Cellulase activity did not increase during hypoxia or N starvation in the presence of aminoethoxyvinylglycine, an inhibitor of ACC synthase, but this inhibition of cellulase induction was reversed during simultaneous exposure to exogenous ethylene. Together these results indicate both the role of ethylene in signaling cell lysis in response to two distinct environmental factors and the significance of hypoxia rather than anoxia in stimulation of ethylene biosynthesis in maize roots.

Arbuscular mycorrhizal fungi and organic fertilizer influence photosynthesis, root phosphatase activity, nutrition, and growth of Ipomoea carnea ssp. fistulosa

The effect of arbuscular mycorrhizal fungi (AMF) inoculation and organic slow release fertilizer (OSRF) on photosynthesis, root phosphatase activity, nutrient acquisition, and growth of Ipomoea carnea N. von Jacquin ssp. fistulosa (K. Von Martinus ex J. Choisy) D. Austin (bush morning glory) was determined in a greenhouse study. The AMF treatments consisted of a commercial isolate of Glomus intraradices and a non-colonized (NonAMF) control. The OSRF was applied at 10, 30, and 100 % of the manufacturer’s recommended rate. AMF plants had a higher net photosynthetic rate (PN), higher leaf elemental N, P, and K, and generally greater growth than NonAMF plants. Total colonization levels of AMF plants ranged from 27 % (100 % OSRF) to 79 % (30 % OSRF). Root acid phosphatase (ACP) and alkaline phosphatase (ALP) activities were generally higher in AMF than non-AMF plants. When compared to NonAMF at 100 % OSRF, AMF plants at 30 % OSRF had higher or comparable ACP and ALP activity, higher leaf elemental P, N, Fe, Cu, and Zn, and a greater PN (at the end of the experiment), leading to generally greater growth parameters with the lower fertility in AMF plants. We suggest that AMF increased nutrient acquisition from an organic fertilizer source by enhancing ACP and ALP activity thus facilitating P acquisition, increasing photosynthesis, and improving plant growth.

Effect of hypobaric conditions on ethylene evolution and growth of lettuce and wheat.

Elevated levels of ethylene occur in enclosed crop production systems and in spaceflight environments, leading to adverse plant growth and sterility. There are engineering advantages in growing plants at hypobaric (reduced atmospheric pressure) conditions in biomass production for extraterrestrial base or spaceflight environments. Objectives of this research were to characterize the influence of hypobaria on growth and ethylene evolution of lettuce (Lactuca sativa) and wheat (Triticum aestivum). Plants were grown under variable total gas pressures [from 30 to 101 kPa (ambient)]. In one study, lettuce and wheat were direct seeded, germinated and grown in the same chambers for 28 d at 50 or 101 kPa. Hypobaria increased plant growth and did not alter germination rate. During a 10-day study, 28-day-old lettuce and 40-day-old wheat seedlings were transplanted together in the same low and ambient pressure chambers; ethylene accumulated in the chambers, but the rate of production by both lettuce and wheat was reduced more than 65% under 30 kPa compared with ambient pressure (101 kPa). Low O2 concentrations [partial pressure of O2 (pO2) = 6.2 kPa] inhibited ethylene production by lettuce under both low (30 kPa) and ambient pressure, whereas ethylene production by wheat was inhibited at low pressure but not low O2 concentration. There was a negative linear correlation between increasing ethylene concentration and decreasing chlorophyll content of lettuce and wheat. Lettuce had higher production of ethylene and showed greater sensitivity to ethylene than wheat. The hypobaric effect on reduced ethylene production was greater than that of just hypoxia (low oxygen).

Separating the effects of hypobaria and hypoxia on lettuce: growth and gas exchange

The objectives of this research were to determine the influence of hypobaria (reduced atmospheric pressure) and reduced partial pressure of oxygen (pO2) [hypoxia] on carbon dioxide (CO2) assimilation (C(A)), dark-period respiration (DPR) and growth of lettuce (Lactuca sativa L. cv. Buttercrunch). Lettuce plants were grown under variable total gas pressures [25 and 101 kPa (ambient)] at 6, 12 or 21 kPa pO2)(approximately the partial pressure in air at normal pressure). Growth of lettuce was comparable between ambient and low total pressure but lower at 6 kPa pO2 (hypoxic) than at 12 or 21 kPa pO2. The specific leaf area of 6 kPa pO2 plants was lower, indicating thicker leaves associated with hypoxia. Roots were most sensitive to hypoxia, with a 50-70% growth reduction. Leaf chlorophyll levels were greater at low than at ambient pressure. Hypobaria and hypoxia did not affect plant water relations. While hypobaria did not adversely affect plant growth or C(A), hypoxia did. There was comparable C(A) and a lower DPR in low than in ambient total pressure plants under non-limiting CO2 levels (100 Pa pCO2, nearly three-fold that in normal air). The C(A)/DPR ratio was higher at low than at ambient total pressure, particularly at 6 kPa pO2- indicating a greater efficiency of C(A)/DPR in low-pressure plants. There was generally no significant interaction between hypoxia and hypobaria. We conclude that lettuce can be grown under subambient pressure ( congruent with25% of normal earth ambient total pressure) without adverse effects on plant growth or gas exchange. Furthermore, hypobaric plants were more resistant to hypoxic conditions that reduced gas exchange and plant growth.

Ethylene reduces gas exchange and growth of lettuce plants under hypobaric and normal atmospheric conditions

Elevated levels of ethylene occur in controlled environment agriculture and in spaceflight environments, leading to adverse plant growth and sterility. The objectives of this research were to characterize the influence of ethylene on carbon dioxide (CO(2)) assimilation (C(A)), dark period respiration (DPR) and growth of lettuce (Lactuca sativa L. cv. Buttercrunch) under ambient and low total pressure conditions. Lettuce plants were grown under variable total gas pressures of 25 kPa (hypobaric) and 101 kPa (ambient) pressure. Endogenously produced ethylene accumulated and reduced C(A), DPR and plant growth of ambient and hypobaric plants. There was a negative linear correlation between increasing ethylene concentrations [from 0 to around 1000 nmol mol(-1) (ppb)] on C(A), DPR and growth of ambient and hypobaric plants. Declines in C(A) and DPR occurred with both exogenous and endogenous ethylene treatments. C(A) was more sensitive to increasing ethylene concentration than DPR. There was a direct, negative effect of increasing ethylene concentration reducing gas exchange as well as an indirect ethylene effect on leaf epinasty, which reduced light capture and C(A). While the C(A) was comparable, there was a lower DPR in hypobaric than ambient pressure plants - independent of ethylene and under non-limiting CO(2) levels (100 Pa pCO(2), nearly three-fold that in normal air). This research shows that lettuce can be grown under hypobaria ( congruent with25% of normal earth ambient total pressure); however, hypobaria caused no significant reduction of endogenous ethylene production.

Ethylene reduces plant gas exchange and growth of lettuce grown from seed to harvest under hypobaric and ambient total pressure

Naturally occurring high levels of ethylene can be a problem in spaceflight and controlled environment agriculture (CEA) leading to sterility and irregular plant growth. There are engineering and safety advantages of growing plants under hypobaria (low pressure) for space habitation. The goals of this research were to successfully grow lettuce (Lactuca sativa cv. Buttercrunch) in a long-term study from seed to harvest under hypobaric conditions, and to investigate how endogenously produced ethylene affects gas exchange and plant growth from seed germination to harvest under hypobaric and ambient total pressure conditions. Lettuce was grown under two levels of total gas pressure [hypobaric or ambient (25 or 101 kPa)] in a long-term, 32-day study. Significant levels of endogenous ethylene occurred by day-15 causing reductions in photosynthesis, dark-period respiration, and a subsequent decrease in plant growth. Hypobaria did not mitigate the adverse ethylene effects on plant growth. Seed germination was not adversely affected by hypobaria, but was reduced by hypoxia (6 kPa pO(2)). Under hypoxia, seed germination was higher under hypobaria than ambient total pressure. This research shows that lettuce can be grown from seed to harvest under hypobaria (≅25% of normal earth ambient total pressure).

Hypobaria and hypoxia affects phytochemical production, gas exchange, and growth of lettuce

Hypobaria (low total atmospheric pressure) is essential in sustainable, energy-efficient plant production systems for long-term space exploration and human habitation on the Moon and Mars. There are also critical engineering, safety, and materials handling advantages of growing plants under hypobaria, including reduced atmospheric leakage from extraterrestrial base environments. The potential for producing crops under hypobaria and manipulating hypoxia (low oxygen stress) to increase health-promoting bioactive compounds is not well characterized. Here we showed that hypobaric-grown lettuce plants (25 kPa ≈ 25% of normal pressure) exposed to hypoxia (6 kPa pO2 ≈ 29% of normal pO2) during the final 3 d of the production cycle had enhanced antioxidant activity, increased synthesis of anthocyananins, phenolics, and carotenoids without reduction of photosynthesis or plant biomass. Net photosynthetic rate (PN) was not affected by total pressure. However, 10 d of hypoxia reduced PN, dark respiration rate (RD), PN/RD ratio, and plant biomass. Growing plants under hypobaria and manipulating hypoxia during crop production to enhance health-promoting bioactive compounds is important for the health and well-being of astronauts exposed to space radiation and other stresses during long-term habitation.

Regulation Of Ethylene Synthesis In Maize Root Responses To Stress

The role and regulation of ethylene synthesis was investigated in maize roots subjected to stresses that inhibit elongation, promote radial swelling and promote formation of aerenchyma. Physical impedance was imposed by compression of the growing medium around unemerged seedlings at controlled pressures, and ethylene production rates by intact seedlings were assayed with the aid of a continuous flow system. Ethylene production increased before effects on growth were observed, and AVG plus STS restored root extension to 90% of control values. One hour after application of 100 kPa pressure to the medium, ACC and conjugated ACC levels and ACC synthase and ACC oxygenase activities all had increased sharply. Transient -N treatment increased sensitivity to ethylene and initiated formation of aerenchyma. Effects of both -N and hypoxia treatments were blocked with Ag+. Both treatments induced synthesis of cellulase which was prevented by AVG. Perception of impedance stress and effects of -N on ethylene binding capacity and affinity are being studied.