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Quality assurance procedures for environmental control and monitoring in plant growth facilities. Report of the North Central Regional 101 Committee on Growth Chamber Use

This report includes procedures for ensuring the quality of the environment provided for plant growth in controlled environment facilities. Biologists and engineers may use these procedures for ensuring quality control during experiments or for ensuring quality control in the design of plant growth facilities. Environmental monitoring prior to and during experiments is included in these procedures. Specific recommendations cover control, acquisition, and calibration for sensor types for the separate parameters of radiation (light), temperature, humidity, carbon dioxide, and air movement.

NASA Discipline Number 61-10

Elimination of toxicity from polyurethane foam plugs used for plant culture

Polyurethane foam plugs commonly are used as collars or supports to grow plants in solution culture. Despite their utility, these foam plugs can be quite toxic to plants, particularly to small seedlings. We have observed tissue injury in tests using plugs to support lettuce, red beet, and potato plants in solution culture. Typically, the injury is initiated on the hypocotyl or stem tissue in direct contact with the foam, and appears within 30 hr as a brownish discoloration on the tissue surface. This discoloration can be followed by complete collapse of affected tissue and eventual death of the seedling. When injury does not progress beyond surface browning, the seedling survives but growth is slowed. In this paper, we report on different treatments that can be used to remove the toxicity of these plugs so they can be used in plant research.

Non-NASA Center

A nondestructive method for continuously monitoring plant growth

In the past, plant growth generally has been measured using destructive methods. This paper describes a nondestructive technique for continuously monitoring plant growth. The technique provides a means of directly and accurately measuring plant growth over both short and long time intervals. Application of this technique to the direct measurement of plant growth rates is illustrated using corn (Zea mays L.) as an example.

NASA Program CELSS

Activity in the ferret: oestradiol effects and circadian rhythms

The present study was conducted to determine whether oestradiol increases activity in the European ferret (Mustela furo), whether this effect is sexually dimorphic, and whether a 24-h rhythm is present in the ferret's daily activity. The activity of male and female adult, postpubertally gonadectomized ferrets was monitored while they were maintained singly on a 13:11 light-dark cycle, before and after implantation with oestradiol-17 beta. Gonadectomized male and female ferrets exhibited equal levels of activity, and neither sex exhibited a significant change in activity following oestradiol implantation. None of the ferrets exhibited a strong circadian rhythm, although weak 24-h rhythms and shorter harmonic rhythms were present. Golden hamsters (Mesocricetus auratus), monitored in an identical manner, exhibited strong circadian rhythms. It was concluded that oestradiol administration may not cause an increase in activity in the ferret, and that this species lacks a strong circadian activity rhythm.

Non-NASA Center

Geoperception in primary and lateral roots of Phaseolus vulgaris (Fabaceae). III. A model to explain the differential georesponsiveness of primary and lateral roots

Half-tipped primary and lateral roots of Phaseolus vulgaris bend toward the side of the root on which the intact half tip remains. Therefore, tips of lateral and primary roots produce growth effectors capable of inducing gravicurvature. The asymmetrical placement of a tip of a lateral root onto a detipped primary root results in the root bending toward the side of the root onto which the tip was placed. That is, the lesser graviresponsiveness of lateral roots as compared with primary roots is not due to the inability of their caps to produce growth inhibitors. The more pronounced graviresponsiveness of primary roots is positively correlated with the presence of columella tissues that are 3.8 times longer, 1.7 times wider, and 10.5 times more voluminous than the columellas of lateral roots. We propose that the lack of graviresponsiveness exhibited by lateral roots is due to the fact that they (i) produce smaller amounts of the inhibitor than primary (i.e., strongly graviresponsive) roots and (ii) are unable to redistribute the inhibitor so as to be able to create a concentration gradient sufficient to induce a pronounced gravitropic response.

NASA Program Space Biology

Correlations between polyamine ratios and growth patterns in seedling roots

The levels of putrescine, cadaverine, spermidine and spermine were determined in seedling roots of pea, tomato, millet and corn, as well as in corn coleoptiles and pea internodes. In all roots, putrescine content increased as elongation progressed, and the putrescine/spermine ratio closely paralleled the sigmoid growth curve up until the time of lateral root initiation. Spermidine and spermine were most abundant near the apices and declined progressively with increasing age of the cells. In the zone of differentiation of root hairs in pea roots, putrescine rose progressively with increasing age, while cadaverine declined. In both pea internodes and corn coleoptiles, the putrescine/spermidine ratio rises with increasing age and elongation. Thus, a block in the conversion of the diamine putrescine to the triamine spermidine may be an important step in the change from cell division to cell elongation.

Non-NASA Center

Polyamine levels as related to growth, differentiation and senescence in protoplast-derived cultures of Vigna aconitifolia and Avena sativa

We have previously reported that aseptically cultured mesophyll protoplasts of Vigna divide rapidly and regenerate into complete plants, while mesophyll protoplasts of Avena divide only sporadically and senesce rapidly after isolation. We measured polyamine titers in such cultures of Vigna and Avena, to study possible correlations between polyamines and cellular behavior. We also deliberately altered polyamine titer by the use of selective inhibitors of polyamine biosynthesis, noting the effects on internal polyamine titer, cell division activity and regenerative events. In Vigna cultures, levels of free and bound putrescine and spermidine increased dramatically as cell division and differentiation progressed. The increase in bound polyamines was largest in embryoid-forming callus tissue while free polyamine titer was highest in root-forming callus. In Avena cultures, the levels of total polyamines decreased as the protoplast senesced. The presence of the inhibitors alpha-difluoromethyl-arginine (specific inhibitor of arginine decarboxylase), alpha-difluoromethylornithine (specific inhibitor of ornithine decarboxylase) and dicyclohexylamine (inhibitor of spermidine synthase) reduced cell division and organogenesis in Vigna cultures. Addition of low concentration of polyamines to such cultures containing inhibitors or removal of inhibitors from the culture medium restored the progress of growth and differentiation with concomitant increase in polyamine levels.

NASA Program Space Biology

Tomato growth as affected by root-zone temperature and the addition of gibberellic acid and kinetin to nutrient solutions

The effect of root-zone temperature on young tomato plants (Lycopersicon esculentum Mill. cv. Heinz 1350) was evaluated in controlled environments using a recirculating solution culture system. Growth rates were measured at root-zone temperatures of 15 degrees, 20 degrees, 25 degrees, and 30 degrees C in a near optimum foliar environment. Optimum growth occurred at 25 degrees to 30 degrees during the first 4 weeks of growth and 20 degrees to 25 degrees during the 5th and 6th weeks. Growth was severely restricted at 15 degrees. Four concentrations of gibberellic acid (GA3) and kinetin were added to the nutrient solution in a separate trial; root-zone temperature was maintained at 15 degrees and 25 degrees. Addition of 15 micromoles GA3 to solutions increased specific leaf area, total leaf area, and dry weight production of plants in both temperature treatments. GA3-induced growth stimulation was greater at 15 degrees than at 25 degrees. GA3 may promote growth by increasing leaf area, enhancing photosynthesis per unit leaf area, or both. Kinetic was not useful in promoting growth at either temperature.

NASA Program CELSS

Effects of artificial enclosure of young lettuce leaves on tipburn incidence and leaf calcium concentration

The young developing leaves of 20-day-old lettuce plants (Lactuca sativa L. 'Buttercrunch') were enclosed by aluminized polyethylene sheaths to decrease transpiration and reduce Ca transport. The plants were grown in recirculating solution culture system using a modified half-strength Hoagland's solution under cool-white fluorescent lamps with a photosynthetic photon flux of 350 micromoles s-1 m-2 in a 16:8-hr (light:dark) period. Air temperature and humidity were 20 degrees C and 65%, respectively. After 4 days of enclosure, 53% of the inner leavers (leaves one to 3 cm in length) were tipburned. After the same period, less than 1% of the inner leaves on control plants were tipburned. The concentration of Ca in enclosed inner leaves was 0.63 mg g-1 dry weight, compared to 1.48 mg g-1 dry weight in inner leaves that were not enclosed. The Ca concentration in transpiring outer leaves of all plants was 9.9 mg g-1 dry weight. The Mg concentration in enclosed inner leaves was 2.25 mg g-1 dry weight, compared to 2.34 mg g-1 dry weight in inner leaves that were not enclosed. This research documents that enclosures of leaves at the growing point, as would occur with normal head development, is sufficient to create a limiting concentration of Ca in the enclosed tissue and encourage tipburn development.

Non-NASA Center

Enhancement of nitrate uptake and growth of barley seedlings by calcium under saline conditions

The effect of Ca2+ on NO3- assimilation in young barley (Hordeum vulgare L. var CM 72) seedlings in the presence and absence of NaCl was studied. Calcium increased the activity of the NO3- transporter under saline conditions, but had little effect under nonsaline conditions. Calcium decreased the induction period for the NO3- transporter under both saline and nonsaline conditions but had little effect on its apparent Km for NO3- both in the presence and absence of NaCl. The enhancement of NO3- transport by Ca2+ under saline conditions was dependent on the presence of Ca2+ in the uptake solution along with the salt, since Ca2+ had no effect when supplied before or after salinity stress. Although Mn2+ and Mg2+ enhanced NO3- uptake under saline conditions, neither was as effective as Ca2+. In longer studies, increasing the Ca2+ concentration in saline nutrient solutions resulted in increases in NO3- assimilation and seedling growth.

NASA Discipline Number 61-10

Growth reponses of eggplant and soybean seedlings to mechanical stress in greenhouse and outdoor environments

Eggplant (Solanum melongena L. var. esculentum 'Burpee's Black Beauty') and soybean [Glycine max (L.) Merr. 'Wells II'] seedlings were assigned to a greenhouse or a windless or windy outdoor environment. Plants within each environment received either periodic seismic (shaking) or thigmic (flexing or rubbing) treatment, or were left undisturbed. Productivity (dry weight) and dimensional (leaf area and stem length) growth parameters generally were reduced more by mechanical stress in the greenhouse (soybean) or outdoor-windless environment (eggplant) than in the outdoor windy environment. Outdoor exposure enhanced both stem and leaf specific weights, whereas mechanical stress enhanced only leaf specific weight. Although both forms of controlled mechanical stress tended to reduce node and internode diameters of soybean, outdoor exposure increased stem diameter.

NASA Discipline Number 61-10

Analysis of growth patterns during gravitropic curvature in roots of Zea mays by use of a computer-based video digitizer

A computer-based video digitizer system is described which allows automated tracking of markers placed on a plant surface. The system uses customized software to calculate relative growth rates at selected positions along the plant surface and to determine rates of gravitropic curvature based on the changing pattern of distribution of the surface markers. The system was used to study the time course of gravitropic curvature and changes in relative growth rate along the upper and lower surface of horizontally-oriented roots of maize (Zea mays L.). The growing region of the root was found to extend from about 1 mm behind the tip to approximately 6 mm behind the tip. In vertically-oriented roots the relative growth rate was maximal at about 2.5 mm behind the tip and declined smoothly on either side of the maximum. Curvature was initiated approximately 30 min after horizontal orientation with maximal (50 degrees) curvature being attained in 3 h. Analysis of surface extension patterns during the response indicated that curvature results from a reduction in growth rate along both the upper and lower surfaces with stronger reduction along the lower surface.

NASA Discipline Plant Biology

Growth and lignification in seedlings exposed to eight days of microgravity

Four-day-old pine seedlings and mung bean and oat seeds were prepared for flight on the third Space Transport System Mission (STS-3). The seedlings and seeds were planted in six mini-growth chambers (two chambers per species) which were placed in a plant growth unit (PGU). Another set of seedlings and seeds was prepared and placed in another PGU as the 1 g control. The flight PGU was positioned in the orbiter mid-deck locker area about 11 h prior to launch. The pine seedlings and germinating mung bean and oat seeds were exposed to 194 h of microgravity. The PGU was received at a temporary laboratory about 75 min post-landing. Plants were observed, photographed and the atmospheric gases analyzed at the landing site. The plants were then brought to our Houston laboratory where they were measured and analyzed for lignin and protein content and for phenylalanine ammonia-lyase (PAL) and peroxidase activities. Flight seedlings were shorter than the controls in all three species. Twenty-five to 40 per cent of the mung bean and oat roots were growing upward, and the mung beans showed signs of disorientation. Flight mung beans showed a significant reduction in lignin content in comparison to the controls, and PAL and peroxidase activities were reduced in flight pine seedlings. The results generally support the postulate that lignin synthesis is reduced in near-weightlessness and show other interesting findings.

Flight Experiment

Statoconia formation in molluscan statocysts

The gravity sensors of all molluscs phylogenetically below the cephalopods are spherical organs called statocysts. The wall of the sphere contains mechanosensory cells whose sensory cilia project into the lumen of the cyst. The lumen is filled with fluid and dense "stones", the statoconia or statoliths, which sink under the influence of gravity to load, and stimulate, those receptor cells which are at the bottom. The statoconia of Aplysia californica are shown to be calcified about a lamellar arrangement of membranes. Similar lamellar membrane arrangements are seen within the receptor cells, and their possible role in the formation of the statoconia is discussed. SEM of unfixed statoconia reveals plate-like crystallization on their surface. Elemental analysis shows a relatively high Sr content, which is of interest, since others have recently reported that Sr is required in the culture medium of several laboratory reared molluscs in order for the statoconia to develop.

Non-NASA Center

Health significance and occurrence of injured bacteria in drinking water

Enteropathogenic and indicator bacteria become injured in drinking water with exposure to sublethal levels of various biological, chemical and physical factors. One manifestation of this injury is the inability to grow and form colonies on selective media containing surfactants. The resulting underestimation of indicator bacteria can lead to a false estimation of water potability. m-T7 medium was developed specifically for the recovery of injured coliforms (both "total" and fecal) in drinking water. The m-T7 method was used to survey operating drinking water treatment and distribution systems for the presence of injured coliforms that were undetected with currently used media. The mean recovery with m-Endo LES medium was less than 1/100 ml while it ranged between 6 and 68/100ml with m-T7 agar. The majority of samples giving positive results with m-T7 medium yielded no detectable coliforms with m-Endo LES agar. Over 95% of the coliform bacteria in these samples were injured. Laboratory experiments were also done to ascribe the virulence of injured waterborne pathogens. Enteropathogens including Salmonella typhimurium, Yersinia enterocolitica and Shigella spp. required up to 20 times the chlorine levels to produce the same injury in enterotoxigenic Escherichia coli (ETEC) and nonpathogenic coliforms. Similar results were seen with Y. enterocolitica exposed to copper. The recovery of ETEC was followed by delayed enterotoxin production, both in vitro and in the gut of experimental animals. This indicates that injured waterborne enteropathogenic bacteria can be virulent.

NASA Discipline Number 13-40

Light-induced import of the chromoprotein, phytochrome, into mitochondria

Mitochondria extracted from plants that were irradiated with actinic light in vivo have associated with them the chromoprotein, phytochrome. This phytochrome retains its native subunit size of 124 kDa after proteolytic treatment of the mitochondria with trypsin and chymotrypsin. This result suggests that phytochrome is not exposed on the outer surface of the outer mitochondrial membrane. Phytochrome, so protected, is not found to be associated with mitochondria derived from unirradiated plants. The possibility that the photoactivation of phytochrome induces a conformational change in its structure which facilitates its transport into the mitochondrion is discussed.

NASA Discipline Plant Biology

Shoot inversion-induced ethylene in Pharbitis nil induces the release of apical dominance by restricting shoot elongation

Shoot inversion induces outgrowth of the highest lateral bud (HLB) adjacent to the bend in the stem in Pharbitis nil. In order to determine whether or not ethylene produced by shoot inversion plays a direct role in promoting or inhibiting bud outgrowth, comparisons were made of endogenous levels of ethylene in the HLB and HLB node of plants with and without inverted shoots. That no changes were found suggests that the control of apical dominance does not involve the direction action of ethylene. This conclusion is further supported by evidence that the direct application of ethylene inhibitors or ethrel to inactive or induced lateral buds has no significant effect on bud outgrowth. The hypothesis that ethylene evolved during shoot inversion indirectly promotes the outgrowth of the highest lateral bud (HLB) in restricting terminal bud (TB) growth is found to be supported by the following observations: (1) the restriction of TB growth appears to occur before the beginning of HLB outgrowth; (2) the treatment of the inverted portion of the shoot with AgNO3, an inhibitor of ethylene action, dramatically eliminates both the restriction of TB growth and the promotion of HLB outgrowth which usually accompany shoot inversion; and (3) the treatment of the upper shoot of an upright plant with ethrel mimics shoot inversion by retarding upper shoot growth and inducing outgrowth of the lateral bud basipetal to the treated region.

NASA Discipline Plant Biology