Kansas Agricultural Statistics

Kansas State Board of Agriculture, U.S. Department of Agriculture, 632 SW Van Buren,

Rm. 200, P.O. Box 3534, Topeka, 66601-3534, USA.

Phone: 913-233-2230 T.J. Byram.

KARL continues number one. Karl and improved Karl remained the leading varieties of wheat seeded in Kansas for the 1995 crop, according to Kansas Agricultural Statistics. Accounting for 22.4 % of the state's wheat, Karl declined slightly from a year ago. In the eastern third of the state, Karl was the most popular variety seeded by a wide margin. Karl was also the number one variety in the north central district, slightly surpassing Pioneer 2163. In the western districts, Tam 107 remained the top variety and was the second most seeded variety state-wide, comprising 20.6 % of the acreage. Pioneer 2163 turned out to be the variety of choice in the central and south central districts and, with 17.1 % of the state's acreage, ranked third overall. Pioneer 2163 was the second leading variety in the eastern third of the state and the north central district. Larned ranked fourth overall, accounting for 7.6 % of the state's acreage. Larned was the number two variety in the western districts. AgriPro Tomahawk ranked fifth in the state, with 7.0 % of the wheat acreage. The top five varieties overall for the state in 1995 are unchanged from last year. The sixth most popular variety in 1995, up from tenth position in 1994, is AGSECO 7853, seeded on 3.7 % of the acreage. AgriPro Thunderbird and Victory are ranked seventh and eighth across the state, at 2.6 and 2.2 % of the acreage, respectively. Newton came in the ninth most popular, with 1.6 % of the acreage, and Tam 200 was tenth, accounting for 1.4 %. Table 1. Distribution of Kansas winter wheat varieties, 1995 crop. ___________________________________________________________________________________________ Agricultural Statistics District _____________________________________________________________ Variety NW WC SW NC C SC NE EC SE State ___________________________________________________________________________________________ PERCENT OF SEEDED ACREAGE Karl/Karl 92 5.9 1.1 3.5 23.7 22.2 32.0 51.4 60.3 75.4 22.4 Tam 107 39.8 57.5 41.1 4.8 8.9 3.1 3.1 0.0 0.9 20.6 2163 1.2 1.0 0.9 21.4 31.0 33.2 23.8 17.8 10.4 17.1 Larned 11.0 14.3 20.2 2.0 4.7 2.0 1.0 0.0 0.0 7.6 AgriPro-Tomahawk 4.8 3.0 3.0 17.8 10.2 7.8 6.0 5.9 1.7 7.0 AGSECO-7853 5.1 3.0 2.0 5.0 5.7 3.9 2.2 2.7 1.0 3.7 AgriPro-Thunderbird 3.5 3.0 1.9 4.2 3.0 2.8 2.1 1.0 0.7 2.6 AgriPro-Victory 0.1 0.0 0.9 8.0 3.1 2.0 3.1 2.2 0.9 2.2 Newton 6.0 2.7 2.9 1.1 1.1 0.3 0.1 0.4 0.1 1.6 Tam 200 1.8 2.0 3.7 1.1 0.9 0.4 0.3 0.0 0.0 1.4 2180 0.0 0.0 1.0 0.0 2.0 4.0 0.1 0.2 0.3 1.3 AgriPro-Pecos 0.0 0.1 0.1 1.7 1.4 2.3 1.8 3.0 1.1 1.1 Eagle 1.5 2.9 3.0 0.2 0.3 0.3 0.1 0.0 0.0 1.1 Scout/Scout 66 1.0 0.8 4.3 0.0 0.2 0.1 0.0 0.0 0.0 1.0 Ike 1.8 2.4 1.2 1.2 0.8 0.4 0.1 0.0 0.0 0.9 AgriPro-Laredo 2.1 0.5 1.4 0.8 0.7 0.3 0.0 0.0 0.0 0.8 Arapahoe 5.2 0.2 0.0 1.2 0.0 0.0 1.1 0.1 0.0 0.8 AgriPro-Longhorn 0.5 0.8 2.5 0.1 0.1 0.7 0.0 0.0 0.0 0.7 AgriPro-Sierra 1.0 0.1 1.0 1.4 1.0 0.3 0.5 1.8 0.0 0.7 AgriPro-Abilene 1.1 0.8 0.3 1.4 0.5 0.2 0.5 0.1 0.0 0.6 AgriPro-Mesa 0.5 0.3 0.2 0.4 0.5 1.0 0.0 0.3 0.0 0.4 Triumph/Triumph 64 0.0 0.0 0.0 0.0 0.1 0.7 0.4 0.0 1.7 0.3 Vista 1.9 0.1 0.0 1.0 0.0 0.0 0.0 0.0 0.0 0.3 Tam 202 0.4 0.3 0.4 0.3 0.1 0.0 0.0 0.0 0.2 0.2 AGSECO-7805 0.0 0.3 0.9 0.1 0.0 0.0 0.0 0.1 0.0 0.2 AgriPro-Ogalala 0.1 0.5 1.0 0.1 0.1 0.0 0.0 0.0 0.0 0.2 AgriPro-Hawk 0.2 0.0 0.7 0.1 0.1 0.2 0.0 0.0 0.0 0.2 Other Hard Var. 3.5 2.3 1.9 0.9 1.3 2.0 2.1 3.3 1.9 2.9 Other Soft Var. 0.0 0.0 0.0 0.0 0.0 0.0 0.2 0.8 3.7 0.1 TOTAL 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 ____________________________________________________________________________________________ Table 2. Distribution of Kansas winter wheat varieties, specified years*. _________________________________________________________________________________________ Variety 1985 1986 1988 1989 1990 1991 1992 1993 1994 1995 _________________________________________________________________________________________ PERCENT OF SEEDED ACREAGE Karl/Karl 92 - - - - 0.7 5.9 11.5 23.0 23.6 22.4 Tam 107 - 0.3 4.9 9.5 14.7 15.4 18.3 19.8 19.0 20.6 2163 - - - - 0.8 2.6 4.6 9.0 13.8 17.1 Larned 8.6 7.9 10.9 9.7 10.7 11.6 8.9 8.3 8.3 7.6 AgriPro-Tomahawk - - - - - - - 1.5 6.2 7.0 AGSECO 7853 - - - - - - 0.2 1.4 2.1 3.7 AgriPro-Thunderbird - - 1.6 7.3 9.3 9.0 7.5 5.5 3.4 2.6 AgriPro-Victory - 0.1 6.2 8.2 7.7 8.2 10.2 8.1 3.9 2.2 Newton 25.7 21.1 13.4 11.6 8.3 7.6 5.8 3.1 2.5 1.6 Tam 200 - - - 0.9 6.1 5.9 4.5 3.2 2.2 1.4 2180 - - - - 0.5 1.6 1.5 1.7 1.4 1.3 AgriPro-Pecos - - - - - - - - 0.2 1.1 Eagle 4.0 3.1 1.4 1.8 1.6 1.1 1.6 1.0 1.1 1.1 Scout/Scout 66 3.6 2.6 2.9 1.8 1.9 1.6 1.8 1.3 1.3 1.0 Ike - - - - - - - - - 0.9 AgriPro-Laredo - - - - - - - - - 0.8 Arapahoe - - - - 0.1 0.1 0.3 0.2 0.8 0.8 AgriPro-Longhorn - - - - - - - - 0.6 0.7 AgriPro-Sierra - - - - - 0.5 1.8 1.4 1.4 0.7 AgriPro-Abilene - - - - 0.6 3.9 4.7 2.2 1.1 0.6 AgriPro-Mesa - - - 1.3 3.4 3.0 1.8 1.2 0.6 0.4 Triumph/Triumph 64 0.6 0.5 0.8 1.0 0.9 0.5 0.5 0.3 0.3 0.3 Vista - - - - - - - - - 0.3 AgriPro-Hawk 12.3 13.5 7.6 4.5 2.9 2.1 0.8 0.7 0.5 0.2 AgriPro-Ogalala - - - - - - - - - 0.2 AGSECO 7805 - - - - 0.1 - 0.2 0.2 0.2 0.2 Tam 202 - - - - - - - - - 0.2 Arkan 6.3 10.1 14.9 11.9 6.8 3.2 2.2 0.8 0.4 0.1 Tam 105 13.4 6.8 1.5 1.0 0.9 1.1 1.0 0.6 0.6 - Other Hard Var. 25.2 33.8 33.8 29.1 21.3 14.4 10.1 5.2 4.1 2.8 Other Soft Var. 0.3 0.2 0.1 0.4 0.7 0.7 0.2 0.3 0.4 0.1 TOTAL 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 100.0 _________________________________________________________________________________________ * No survey done for the 1987 crop.

Kansas State University

Department of Agronomy, Throckmorton Hall, Manhattan, KS 66506, USA.

Plant regeneration from protoplast-derived haploid embryogenic callus of wheat (Triticum aestivum). X. Gu and G.H. Liang. Plant regeneration from wheat protoplasts has been successful in recent years. However, most of these studies used cell suspension cultures as the sources of protoplast culture. Only in two instances were protoplasts directly obtained from embryogenic calli that were derived from immature embryos or from the inflorescence. Because protoplasts can take up exogenous DNA efficiently when they are treated with electroporation or PEG, it is desirable to establish a procedure whereby protoplast isolation and culture is dependable, stable, efficient, and requires less time. We report herewith the direct isolation of protoplasts from embryogenic calli derived from anthers and regeneration of green plants from protocalli in two wheat cultivars, without using cell suspension cultures. Spikes of winter and spring wheat cultivars (Jinghua, Karl 92, and Pavon 76) were collected from plants grown in a greenhouse or a growth chamber, and anthers containing mid- and late-uninucleated microspores were aseptically excised and placed on N6 and W14 media, respectively, to induce callus at 26-28 C in darkness. Calli were produced 1 month later, and embryogenic calli were identified and transferred to maintenance medium (MS) supplemented with 2 mg/l 2,4-D and 3 % sucrose. Cultures were incubated at 25-28 C in the dark and rigorous selection for embryogenic calli was carried out in every subculture cycle (3 to 4 weeks). Embryogenic calli were transferred to a fresh medium for 1 to 2 weeks to prepare protoplasts for isolation. About 1-2 g of embryogenic calli were suspended in an enzyme solution, and the mixture was placed on a rotary shaker (ca 80 rpm) for 2-4 h and kept in a stationary condition for another 5-7 h at 28 C in the dark. The isolated protoplasts were filtered, washed, purified, and then resuspended in culture medium with a density of 1-5 x 10⁵/ml adjusted by using a hemacytometer. Protoplast-derived calli larger than 1 mm were transferred to a solid medium (MS) for continued development. After 3 weeks, the embryogenic calli with organized structures were selected and transferred to a MS differentiation medium and grown at 24-26 C in a 16/8 h light-dark condition to promote shoot induction. Plantlets were transferred to a hormone-free half strength MS medium or a MS medium with 1-0.5 mg/l IAA for root development. The frequency of embryogenic calli from the three cultivars ranged from 0.00 to 3.19 % on W14 medium and 1.10 to 4.29 % on N6 medium. The average frequencies of callus induction and embryogenic callus induction were not significantly different between Jinghua and Pavon 76, but they were higher than that of Karl 92. Isolation of protoplasts using an enzyme solution was enhanced by shaking in the initial stage, and damage of the protoplasts was reduced by keeping the mixture in a stationary condition following shaking. Embryogenic calli at an age of 8-11 days of subculture produced the maximum yield of protoplasts (6.0-8.0 x 10⁵/ml). A new cell wall was regenerated from 1 to 3 days after incubation, first cell divisions were observed at 2-4 days of culture, and the second and third divisions were seen within 1-2 weeks. The frequencies of protoplast division were different among different densities of initial culture. Cells, at a density of 3-4 x 10⁵/ml showed the highest frequency of division. No cell division or few cell divisions were observed in densities less than 1 x 10⁵/ml. Addition of fresh medium was not necessary during protoplast culture. If more than 0.5 ml fresh medium with a relatively higher concentration of glucose (0.3 M to 0.5 M) was added during the first 14 days, cells elongated and the cytoplasm shrank, but if less than 0.5 ml of fresh medium without glucose was added, the cells multiplied and protocalli formed. After culture for 1 month, protoplasts from Jinghua and Pavon 76 were able to form microcalli on both modified PMI and KM8p media, but protocallus formation was relatively higher for Pavon 76 than for Jinghua. The frequencies of colony formation varied from 0.2 % to 0.5 % for Jinghua and from 0.1 % to 2 % for Pavon 76. The cell division of Karl 92 was very low, and no protocalli formed on either PMI or KM8p media, although the embryogenic calli of Karl 92 were friable and grew faster on the maintenance medium. The protocolonies of Jinghua were transferred to a solid medium for continued growth. Some microcalli turned brown and watery, but others became undifferentiated, with a hard texture. The organized structures and embryoids were transferred to a differentiation medium, and green shoots formed after 3-4 weeks. More than 10 green plants of Jinghua from protoplast culture were transferred to vermiculite. The protocalli of Pavon 76 grew fast, and after 2 to 4 weeks of subculture, some microcalli became undifferentiated and friable type of calli. The protoplast-derived embryogenic calli of Pavon 76 formed about 15 to 20 small green plantlets and four green plants on differentiation medium and then on rooting medium.

Direct gene transfer through microprojectile bombardment of immature embryos and regenerable embryogenic callus of wheat. X. Gu, S. Muthukrishnan, and G.H. Liang. Transgenic plants have been obtained recently from wheat through high-velocity bombardment of plant cells with DNA-coated gold (or tungsten) particles. In most of these cases, the Bar gene was used as a selectable marker, and herbicide-resistant transgenic plants were obtained. However, useful transgenic plants that are insect-, virus-, and fungus-resistant are still lacking in wheat. The hygromycin phosphotransferase (hph) gene that confers resistance to hygromycin was used to obtain stably transformed cell lines of Triticum monococcum. However, hygromycin as a selectable marker is not used successfully in common wheat (T. Aestivum), although the hph gene is used widely for the production of transgenic rice and some other grasses. In order to obtain transgenic wheat plants for insect resistance, we used plasmid DNA containing a chitinase gene and the hph gene for direct gene transfer. Plant Material and Tissue Culture. Wheat plants (T. aestivum cvs. Pavon 76, Hartog, Chinese Spring, Karl 92, and Jinghua No.1) are grown in a greenhouse. To establish callus cultures, immature embryos 12 to 14 d postanthesis are surface sterilized with 70 % ethanol for 2 min and 1.05 % sodium hypochlorite for 15 min, followed by three washes with sterile distilled water. Immature embryos are placed aseptically with the scutellum exposed on MS medium with 2 mg/l 2,4-D and 3 % sucrose. Twenty- five embryos are placed in each petri dish (100 x 15 mm). Embryogenic calli are maintained at 25-27 C on the same MS medium and subcultured at 3- to 4-week intervals. Plasmid DNA. The PGL2CaMVG11 gene from rice and the PGL2CaMVMSEC gene from tobacco hornworm are kindly provided by W. Lin and S. Muthukrishnan (Department of Biochemistry, KSU). The vector, PGL2, consists of a marker gene, hph, conferring hygromycin resistance and encoding a rice chitinase gene, G11, and the Manduca sexta endochitinase gene under the control of the cauliflower mosaic virus (CaMV) 35S promoter. Microprojectile Bombardment and Selection of Resistant Lines. Prior to bombardment, plasmid DNAs are precipitated and adsorbed to gold (1 micro m) or M17 tungsten particles following the procedure recommended by the manufacturer (Bio-Rad) and Weeks et al. DNA-coated particles are delivered to target tissues using PDS-1000/He device. For every bombardment, 10 micro l of the DNA-gold (or tungsten) particle suspension is placed in the center of the microcarrier. Twenty-five immature embryos are placed in the center of a 100 x 15 mm Petri dish containing MS medium with 2 mg/l 2,4-D and 3 % sucrose. After 5 days of culture, the embryo-derived calli are bombarded. The distance from the stopping screen to the target is 10-13 cm, and the rupture disc strength is 900 and 1,100 p.s.i. To evaluate the effect of osmolarity in the culture medium, 0.4 M mannitol is added, the embryos are maintained on a mannitol-containing medium for 4 h before and 16 h after the bombardment, and then transferred to MS medium with 2 mg/l 2,4-D and 3 % sucrose. After bombardment, embryo-derived calli are maintained on the same medium for 1 week and then transferred to a MS selection medium containing 1 mg/l 2,4-D and 20-30 mg/l of hygromycin (MS1H20 and MS1H30). The calli then are subcultured every 14 days at 24-26 C under a 16/8 light-dark period. Plant regeneration. For regeneration, resistant calli with green spots or green shoots selected on MS1H20 or MS1H30 are transferred to regeneration media (MS medium supplemented with 1 mg/l kinetin, 1-2 mg/l BA, and 20-30 mg/l hygromycin or with 1 mg/l dicamba and 20-30 mg/l hygromycin). All cultures are under 16/8 light-dark at 24-26 C until the plantlets are at least 2-4 cm tall. Green shoots with a well-developed root system are transferred directly to soil and grown in a temperature-and light-controlled growth chamber. The shoots without a well-developed root system are transferred to a rooting medium containing half-strength MS elements and supplemented with 0.5 mg/l NAA, 30 mg/l hygromycin; then the plantlets are transferred to soil pots and acclimated under lower humidity at 18-21 C with 16 h light in a growth chamber. Protein extract and Western blot analysis. Tissues (0.5-1.0 g) from transgenic plants are ground in liquid nitrogen with a small mortar and pestle. Extracts are made with 1.5 ml of 0.1 M KH₂PO₄ (pH 6.5) and 0.5 mM PMSF. Protein concentrations are determined by a modified BCA method (Pierce) with bovine serum albumin (BSA) as a standard. Proteins are precipitated with trichloroacetic acid before dissolving in an SDS-containing buffer and loading into gels. Western blots of denaturing or nondenaturing PAGE are made as described by Sambrook et al. After protein gel electrophoresis, the proteins are transferred to a nitrocellulose membrane. Anti-bean-chitinase rabbit serum is used as the primary antibody to detect chitinase in the membrane. Horseradish peroxidase (HRP) conjugated with goat anti-rabbit IgG(H+L) is used as the secondary antibody. Genomic DNA isolation and Southern blot analysis. Genomic DNA is isolated from leaves of transgenic plants according to the CTAB nucleic acid extraction procedure. DNA is treated with Rnase, and samples of 15 FONTSIZE=2 FACE="WP MathA"Fg of the total DNA are digested with either HindIII or EcoRV in a final volume of 150 micro l. The digested DNA is precipitated by ethanol, and electrophoresed on 0.8 % agarose gels. After electrophoresis, DNA fragments are denatured and transferred to Gene Screen Plus membrane (Du Pont) using the method described by Maniatis et al. The 537-bp ScaI-HindIII DNA fragment of the G11 chitinase gene is labeled by (FONT SIZE=2 FACE="WP Greek Century""-P³²)-DCTP using the BRL Random Primer. A DNA Labeling System is used as a hybridization probe, and the manufacturer's instructions are followed for Southern blot analysis.

Evapotranspiration Laboratory

Department of Agronomy, Kansas State University, Manhattan, KS 66506, USA. M.B. Kirkham.

Ellipsoidal description of water flow into soil under a disc permeameter. Understanding water movement in wheat fields is important. In field soil, we can distinguish between macroporosity and matrix porosity. Macroporosity refers to the interconnected pore space of voids, which causes preferential transport of both water and chemicals. When transport occurs through the macropores, exchange of water between the macropores and the pores of the matrix is limited. Matrix porosity refers to those pores in which the flow through the body of the soil is slow enough so that extensive interpore mixing occurs. Macropores are frequently more extensive near the surface of the soil, because much of the biologically created macroporosity drops off with depth as the populations of soil flora and fauna decline. For proper agricultural water management in wheat fields, characterization of the hydraulic properties of soils in the tillage (surface) layer is paramount. Tillage affects pore size. Pores are smaller in tilled soils, because tillage pulverizes the soil. When the soil is not tilled, decaying roots and other organic matter create voids. Also, earthworms thrive on the organic matter, and their populations are greater in soil that has not been tilled. Recognition of the importance of macropores and preferential flow has led to the development of instruments that can be used in the field to control preferential water flow through macropores and soil cracks. The first practical instrument was developed in 1981 by Brent E. Clothier of New Zealand and Ian White of Australia and is known as the tension infiltrometer, which evolved into the disc permeameter. With these instruments, the amount of macropore flow measured is controlled by applying water to soil at water potentials less than 0. The disc permeameter is being used widely to characterize the hydraulic properties of the tilth layer. Therefore, it is important to describe the multidimensional flow of water away from this circular source. In the past year, Kirkham and Clothier (1994a, 1994b) used an unique idea to analyze the flow pattern, because they assumed that the three-dimensional wetting fronts under the circular source are ellipsoidal. Experimental data showed that assumption was valid, and wet fronts under the disc form ellipsoids. By knowing that the infiltrated water forms the volume of an ellipsoid, one can assess the depth of penetration of water simply by measuring with a ruler on the surface of the soil the distance the water has moved away from the disc permeameter. From this ellipsoidal assessment of the depth of penetration of infiltrated water, the depth over which the disc permeameter provides a measure of the soil's physical properties can be inferred easily. (Acknowledgement: The help of Prof. Don Kirkham, Iowa State University, who suggested the ellipsoidal idea, is gratefully acknowledged.)

Publications. Kirkham MB and Clothier BE. 1994a. Ellipsoidal description of water flow into soil from a surface disc. Transactions of the 15th International Congress of Soil Science 2b:38-39. Kirkham MB and Clothier BE. 1994b. Wetted soil volume under a circular source In: Proc 13th International Conference, International Soil Tillage Research Organization (Jensen HE, Schjonning P, Mikkelsen SA, and Madsen KB eds). The Royal Veterinary and Agricultural University and the Danish Institute of Plant and Soil Science, Lyngby and Copenhagen, Denmark. Pp. 573-578. Kirkham MB. 1994. Soil-water relationships. In: Encyclopedia of Agricultural Science (Arntzen CJ, Ed-in-Chief, and Ritter EM, Assoc Ed). Academic Press, San Diego. 4:151-168. Zhang J and Kirkham MB. 1994. Drought-stress-induced changes in activities of superoxide dismutase, catalase, and peroxidase in wheat species. Plant Cell Phys 35:785-791.

Kansas State University Department of Plant Pathology, Throckmorton Hall, Manhattan, KS 66506-5502, USA. M. Pyle¹, M. Mazzola², P. Harris³, P.W. Stahlman³, and J.E. Lea ch¹. ¹ KSU Department of Plant Pathology, Manhattan, KS; ² USDA-ARS, Pullman, WA; and ³ FHSU Agricultural Experiment Station, Hays, KS 67601, USA.