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Rapid Testing of Salinity Effects on Pistachio Seedling Rootstocks

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Rapid Testing of Salinity Effects on Pistachio Seedling Rootstocks J. AMER. Soc. HORT. SCI. 116(3):555-559. 1991. Rapid Testing of Salinity Effects on Pistachio Seedling Rootstock G.A. Picchionil and S. Miyamoto2 Texes A&M University Agricultural Research and Extension Center, 1380 A&M Circle, El Paso, TX 79927 J.B. Storey3 Department of Horticultural Sciences, Texas A&M University, College Station, TX 77843-2133 Additional index words. sodium chloride, sodium : calcium ratio, excised roots, Pistacia spp., screening methods, saline tolerance, sodium exclusion, membrane permeability Abstract. A laboratory procedure was used to evaluate saline tolerance of pistachio rootstock species. Results were compared to those from a 2-year, outdoor lysimeter study to test reliability of the method. Excised root tips from seedlings of Pistacia atlantica Desf., P. terebinthus L. (two selections), and P. integerrima Stewart × atlantica (Pioneer Gold II, or PG II), were exposed to laboratory solutions that simulated soil solution electrical conductivity (EC) and Na : Ca ratios in the lysimeters. Following 24 hours of incubation, the efflux of ultraviolet (UV)-absorbing solutes was measured, providing an indication of cell membrane permeability. Leakage occurred with saline solutions com- parable to lysimeter soil water salinity that increased leaf Na concentrations and decreased average root growth (175 -l 2 mM NaCl with 12.5 mM Ca, or EC of 18.1 dS· m ). Cell injury increased linearly with salinity (R = 0.81) and was highest in root tips of a P. terebinthus selection having least Na exclusion capability in the lysimeters. On average, these excised roots lost 38% more solutes than roots of a stronger Na-excluding genotype. There were no differences in leakage responses of the other species and selections. Leakage intensity was independent of various stress media, including isosmotic CaC12, mannitol, and the simulated Na/Ca mixtures in molar ratios of 10:1 to 20:1. With no Ca, however, damage caused by isosmotic NaCl was 76% to 87% higher, indicating that for these species, the Na : Ca ratio can alter root cell membrane permeability. Correlation between long-term observations in the lysimeters and leakage occurrence in the laboratory indicates that solute leakage tests with roots may aid in characterizing Pistacia spp. rootstocks for saline condition. Empirical screening of rootstock for saline resistance can injury is interpreted as direct injury to cell membranes (Leopold involve lengthy and extensive evaluations, but a simpler method and Willing, 1984; Willing and Leopold, 1983). could aid in the characterization process. The reliability of a We could find no reports incorporating long-term observa- rapid procedure could be determined by comparing results to tions of growth and ion uptake of salt-treated rootstock with responses obtained in a more natural environment, such as out- short-term laboratory responses using the UV-absorbing solute door lysimeters. For example, Greenway (1970) proposed that technique. Cell membranes of roots must endure immediate ef- short-term studies of excised roots, which reveal immediate con- fects of salinity due to direct exposure to the soil solution (Maas sequences of salinity, may provide useful comparisons to long- and Nieman, 1978), a condition that could be simulated in the term responses measured in natural conditions. Based on results laboratory and observed by UV-absorption measurement. with various crop species,. Nassery (1979) suggested that mea- The objectives of this study were to 1) assess salinity re- surement of K loss from excised root segments may accurately sponses of root tips from two pistachio rootstock species and and rapidly estimate salt resistance of plants. Milbocker (1988) one hybrid using the UV-absorbing solute technique, and 2) achieved a close correlation between the induction of cell plas- compare these results to those obtained from a previous, long- molysis in root tips and a conventional, long-term screening term study involving the same rootstock. procedure for the ranking of salt resistance of seven azalea cul- tivars. Materials and Methods Passive release of UV-absorbing solutes from plant tissues Seedling culture and test material. Seedlings of P. atlantica also measures short-term salinity effects. Rauser and Hanson (PI 246336), P. terebinthus (PI 246341 and 246342, referred to (1966) showed Na and K salts (50 mM concentrations and above) as A and B, respectively), and P. integerrima × atlantica (PG induced leakage of UV-absorbing materials from excised root II) were maintained in an open shadehouse for 3 years in 10- tips of soybean. Several workers have applied this criterion to liter plastic pots containing Hueco sandy loam soil (calcareous, the study of salt-affected leaf disks (Leopold and Willing, 1984; coarse silty, mixed, thermic, Petrocalcic Paleargid). The seed- Redmann et al., 1986). Leakage of UV-absorbing solutes upon lings were from the same experimental population of a previous salinity study made in outdoor lysimeters (Picchioni et al., 1990) but had not been used previously. On 22 Mar. 1989, dormant Received for publication 13 Mar. 1990. Contribution from the Texas Agricul- plants were moved to a greenhouse and tops were pruned back tural Experiment Station and supported in part by the Expanded Research Area Fund of the Experiment Station. Appreciation is extended to the Entek Corp. by »25% (to 0.3 to 0.6 m high). Roots were washed free of (Grapevine, Texas) for donation of ‘Micro II” tubing used for solution cul- soil, and individual seedlings were transplanted into sealed, 16- tures. The cost of publishing this paper was defrayed in part by the payment of liter plastic pots containing complete, full-strength Hoagland page charges. Under postal regulations, this paper therefore must be hereby nutrient solution #1 (Hoagland and Arnon, 1950). Aeration was marked advertisement solely to indicate this fact. provided continuously by “Micro II” tubing (Entek Corp., ‘Graduate research assistant and Thomas B. Slick Fellow. Present address: Grapevine, Texas), glued to the base of each pot, and connected Dept. of Pomology, Univ. of California, Davis, CA 95616. -4 3 2Professor of Soil and Water Sciences. to a pressure pump providing a flow rate of 3.2 × 10 m /sec 3Professor of Horticultural Sciences. per pot. The greenhouse was maintained between a minimum J. Amer. Soc. Hort. Sci. 116(3):555-559. 1991. 555 of 21C at night and maximum of 27C during the day. Through- content in the lysimeter soils (Picchioni et al., 1990) were ap- out the 7-week study, nutrient solution pH varied between 5.5 plied to P. atlantica, P. terebinthus A, and PG II roots. The and 6.5, and was renewed and adjusted to pH 5.5 (with 1 N EC values for the control, 125:12.5, 175:17.5, and 250:25.0 -3 7, , NaOH or HC1) every 14 days. Midway between changes, so- (mM Na : Ca) treatment solutions were 2.3 × 10 ,14. 18.7 lution was added to replace transpirational losses (ranging from and 24.9 dS·m-1, respectively. To more closely simulate lysi- -1 150 to 300 ml·day ) and maintain upper structural roots below meter soils, which had a CaSO4 volubility limit in solution (Pic- the surface of the solution. chioni et al., 1990), Ca concentrations were held constant in Within 4 weeks, seedlings had vigorous tops with fully ex- two additional solutions at the highest Na concentrations, pro- panded leaves, three to five elongating shoots, and new roots. viding increased Na : Ca ratios (14:1 and 20:1). Salinity of the Such roots are described for other woody species as having rapid additional 175:12.5 and 250:12.5 solutions was 17.4 and 22.3 extension growth and subsequent secondary thickening; e.g., dS·m-1, respectively. white “extension” roots of grapevine (Richards, 1983), and Responses of P. terebinthus genotypes (trial 2). The leakage white “axial” roots of various rootstock (Rem, 1987). They from roots of P. terebinthus A and B selections was compared are likely an important zone for water and nutrient absorption. in duplicate trials, and the results were combined. P. terebinthus In our study, visible laterals (³5 cm proximal to the tip) were B seedlings have been shown to maintain the lowest root and formed no sooner than 3 to 4 weeks after appearance of the basal stem but highest leaf Na concentrations at high salinity primary structures. In soil environments, new roots of P. inte- compared to other rootstock selections (Picchioni et al., 1990). gerrima have similar characteristics (e.g., white tips with rapid Only the control, 175:12.5, and 250:12.5 treatments or those elongation) when observed through rhizotrons (Goldhammer et of equivalent osmolality were applied in this and in subsequent al., 1989; D. Goldhammer, personal communication). With the trials. The EC values were 2.5 × 10-2, 17.6, and 23.2 dS·m-l, exception of diameter (0.5 to 1.5 mm), selected roots were respectively. morphologically similar. Isosmotic solutions (trials 3 and 4). To evaluate solute spec- Testing procedure. Four solute leakage trials (1 to 4) were ificity, Na/Ca, CaC12, mannitol, and NaCl without Ca (Na/0 made between 21 Apr. and 6 June 1989. All laboratory proce- Ca) were applied to PG II (trial 3) and P. atlantica (trial 4). dures were developed in preliminary experiments (Picchioni, The treatments were isosmotic to the 175:12.5 and 250:12.5 -1 1989). One root per plant per treatment was removed in the Na/Ca solutions (359 and 497 mOsm·kg , respectively). Os- greenhouse, then shortened to 2 cm from the tip using a razor molality, measured with a cryoscopic osmometer (Osmette S, blade. Preliminary observation indicated that the distal 1 to 2 Precision Instruments, Sudbury, Mass.), varied by <0.3% among cm was the most salt-sensitive zone of new roots. Root tips the isosmotic solutions.
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