{ "document_id": 15942, "page": 2, "width_pt": 684, "height_pt": 864, "rotation": 0, "pipeline_version": "paddleocr-vl-1.6+paul-48", "engine": "paddleocr_vl", "source_run_id": 16053, "item_count": 38, "counts_by_type": { "caption": 1, "footer": 2, "heading": 1, "table": 1, "table_cell": 26, "text": 7 }, "build_ms": 22, "items": [ { "id": "p2.1", "order": 1, "type": "caption", "bbox": { "l": 79.68, "t": 777.36, "r": 333.6, "b": 766.56 }, "text": "Table 1. Genetic background of 19 pawpaw (Asimina triloba) selections.", "attrs": { "block_label": "figure_title" } }, { "id": "p2.2", "order": 2, "type": "table", "bbox": { "l": 79.92, "t": 766.56, "r": 332.64, "b": 488.64 }, "text": "", "attrs": { "block_label": "table", "n_rows": 13, "n_cols": 2, "integrity": { "cells_checked": 26, "cells_flagged": 5, "flags": [ { "row": 1, "col": 1, "coverage": 0, "text": "Wild seedling from Hart County, Ky." }, { "row": 2, "col": 1, "coverage": 0.27, "text": "Seedling of ‘Overleese’ female × ‘Davis’ male selected in E…" }, { "row": 4, "col": 1, "coverage": 0.13, "text": "‘Davis’ female × ‘Overleese’ male selected in Ontario, Cana…" }, { "row": 6, "col": 0, "coverage": 0.5, "text": "10) Prolific" }, { "row": 10, "col": 0, "coverage": 0.5, "text": "14) Sunflower" } ] } }, "children": [ { "id": "p2.2.1", "order": 3, "type": "table_cell", "bbox": { "l": 82.50108, "t": 762.5004, "r": 135.98808, "b": 731.5008 }, "text": "Cultivar\n1) Cales Creek\n2) Davis", "attrs": { "row": 0, "col": 0, "rowspan": 1, "colspan": 1, "bbox_refit": true, "merged_continuation": true } }, { "id": "p2.2.2", "order": 4, "type": "table_cell", "bbox": { "l": 154.50108, "t": 762.5004, "r": 303.48708, "b": 731.5008 }, "text": "Genetic background\nWild seedling from Summers Co., W. Va.\nWild seedling from Eaton Rapids, Mich.", "attrs": { "row": 0, "col": 1, "rowspan": 1, "colspan": 1, "bbox_refit": true, "merged_continuation": true } }, { "id": "p2.2.3", "order": 5, "type": "table_cell", "bbox": { "l": 82.50108, "t": 729.501, "r": 152.47608, "b": 720.501 }, "text": "3) Greenriver Belle", "attrs": { "row": 1, "col": 0, "rowspan": 1, "colspan": 1, "bbox_refit": true } }, { "id": "p2.2.4", "order": 6, "type": "table_cell", "bbox": { "l": 206.28, "t": 724.872, "r": 332.64, "b": 710.976 }, "text": "Wild seedling from Hart County, Ky.", "attrs": { "row": 1, "col": 1, "rowspan": 1, "colspan": 1 } }, { "id": "p2.2.5", "order": 7, "type": "table_cell", "bbox": { "l": 82.50108, "t": 718.5012, "r": 107.24208, "b": 709.5012 }, "text": "4) IXL", "attrs": { "row": 2, "col": 0, "rowspan": 1, "colspan": 1, "bbox_refit": true } }, { "id": "p2.2.6", "order": 8, "type": "table_cell", "bbox": { "l": 206.28, "t": 710.976, "r": 332.64, "b": 697.08 }, "text": "Seedling of ‘Overleese’ female × ‘Davis’ male selected in Eaton Rapids, Mich.", "attrs": { "row": 2, "col": 1, "rowspan": 1, "colspan": 1 } }, { "id": "p2.2.7", "order": 9, "type": "table_cell", "bbox": { "l": 82.50009, "t": 697.5015, "r": 136.24809, "b": 677.5017 }, "text": "5) Middletown\n6) Mitchell", "attrs": { "row": 3, "col": 0, "rowspan": 1, "colspan": 1, "bbox_refit": true, "merged_continuation": true } }, { "id": "p2.2.8", "order": 10, "type": "table_cell", "bbox": { "l": 154.50009, "t": 697.5015, "r": 295.13409, "b": 677.5017 }, "text": "Wild seedling from Middletown, Ohio.\nWild seedling from Iuka, Ill.", "attrs": { "row": 3, "col": 1, "rowspan": 1, "colspan": 1, "bbox_refit": true, "merged_continuation": true } }, { "id": "p2.2.9", "order": 11, "type": "table_cell", "bbox": { "l": 82.50009, "t": 675.5019, "r": 112.24509, "b": 666.5019 }, "text": "7) NC-1", "attrs": { "row": 4, "col": 0, "rowspan": 1, "colspan": 1, "bbox_refit": true } }, { "id": "p2.2.10", "order": 12, "type": "table_cell", "bbox": { "l": 206.28, "t": 669.288, "r": 332.64, "b": 655.392 }, "text": "‘Davis’ female × ‘Overleese’ male selected in Ontario, Canada.", "attrs": { "row": 4, "col": 1, "rowspan": 1, "colspan": 1 } }, { "id": "p2.2.11", "order": 13, "type": "table_cell", "bbox": { "l": 82.49991, "t": 653.5023, "r": 147.40791, "b": 633.5025 }, "text": "8) Overleese\n9) PA-Golden(#1)", "attrs": { "row": 5, "col": 0, "rowspan": 1, "colspan": 1, "bbox_refit": true, "merged_continuation": true } }, { "id": "p2.2.12", "order": 14, "type": "table_cell", "bbox": { "l": 154.49991, "t": 653.5023, "r": 309.98391, "b": 622.5027 }, "text": "Seedling from Rushville, Ind.\nSeedling from G.A. Zimmerman collection selected in Amherst, N.Y.", "attrs": { "row": 5, "col": 1, "rowspan": 1, "colspan": 1, "bbox_refit": true, "merged_continuation": true } }, { "id": "p2.2.13", "order": 15, "type": "table_cell", "bbox": { "l": 79.92, "t": 627.6, "r": 206.28, "b": 613.704 }, "text": "10) Prolific", "attrs": { "row": 6, "col": 0, "rowspan": 1, "colspan": 1 } }, { "id": "p2.2.14", "order": 16, "type": "table_cell", "bbox": { "l": 154.50009, "t": 620.5029, "r": 281.99409, "b": 611.5029 }, "text": "Seedling from Eaton Rapids, Mich.", "attrs": { "row": 6, "col": 1, "rowspan": 1, "colspan": 1, "bbox_refit": true } }, { "id": "p2.2.15", "order": 17, "type": "table_cell", "bbox": { "l": 79.92, "t": 613.704, "r": 206.28, "b": 599.808 }, "text": "11) Rebecca’s Gold", "attrs": { "row": 7, "col": 0, "rowspan": 1, "colspan": 1 } }, { "id": "p2.2.16", "order": 18, "type": "table_cell", "bbox": { "l": 206.28, "t": 613.704, "r": 332.64, "b": 599.808 }, "text": "Seedling from Eaton Rapids, Mich.", "attrs": { "row": 7, "col": 1, "rowspan": 1, "colspan": 1 } }, { "id": "p2.2.17", "order": 19, "type": "table_cell", "bbox": { "l": 82.5, "t": 598.125, "r": 147.220139, "b": 589.625 }, "text": "12) SAA-Zimmerman", "attrs": { "row": 8, "col": 0, "rowspan": 1, "colspan": 1, "bbox_refit": true } }, { "id": "p2.2.18", "order": 20, "type": "table_cell", "bbox": { "l": 154.50019, "t": 598.5, "r": 309.98419, "b": 578.5002 }, "text": "Seedling from G.A. Zimmerman collection selected in Amherst, N.Y.", "attrs": { "row": 8, "col": 1, "rowspan": 1, "colspan": 1, "bbox_refit": true } }, { "id": "p2.2.19", "order": 21, "type": "table_cell", "bbox": { "l": 82.50019, "t": 576.5004, "r": 110.24719, "b": 567.5004 }, "text": "13) Sue", "attrs": { "row": 9, "col": 0, "rowspan": 1, "colspan": 1, "bbox_refit": true } }, { "id": "p2.2.20", "order": 22, "type": "table_cell", "bbox": { "l": 154.50019, "t": 576.5004, "r": 240.63019, "b": 567.5004 }, "text": "Wild seedling from Ind.", "attrs": { "row": 9, "col": 1, "rowspan": 1, "colspan": 1, "bbox_refit": true } }, { "id": "p2.2.21", "order": 23, "type": "table_cell", "bbox": { "l": 79.92, "t": 572.016, "r": 206.28, "b": 558.12 }, "text": "14) Sunflower", "attrs": { "row": 10, "col": 0, "rowspan": 1, "colspan": 1 } }, { "id": "p2.2.22", "order": 24, "type": "table_cell", "bbox": { "l": 154.50406, "t": 565.5006000000001, "r": 258.63406, "b": 556.5006000000001 }, "text": "Wild seedling from Chanute, Kansas.", "attrs": { "row": 10, "col": 1, "rowspan": 1, "colspan": 1, "bbox_refit": true } }, { "id": "p2.2.23", "order": 25, "type": "table_cell", "bbox": { "l": 82.50136, "t": 554.5008, "r": 139.99336, "b": 512.5014 }, "text": "15) Sweet Alice\n16) Taylor\n17) Taytwo\n18) Wells", "attrs": { "row": 11, "col": 0, "rowspan": 1, "colspan": 1, "bbox_refit": true, "merged_continuation": true } }, { "id": "p2.2.24", "order": 26, "type": "table_cell", "bbox": { "l": 154.50136, "t": 554.5008, "r": 311.48836, "b": 501.5016 }, "text": "Wild seedling from W. Va.\nWild seedling from Eaton Rapids, Mich.\nWild seedling from Eaton Rapids, Mich.\nCultivated seedlings (open pollinated) from Salem, Ind.", "attrs": { "row": 11, "col": 1, "rowspan": 1, "colspan": 1, "bbox_refit": true, "merged_continuation": true } }, { "id": "p2.2.25", "order": 27, "type": "table_cell", "bbox": { "l": 82.50136, "t": 499.5018, "r": 122.22736, "b": 490.5018 }, "text": "19) Wilson", "attrs": { "row": 12, "col": 0, "rowspan": 1, "colspan": 1, "bbox_refit": true } }, { "id": "p2.2.26", "order": 28, "type": "table_cell", "bbox": { "l": 154.49236, "t": 499.5018, "r": 288.03436, "b": 490.5018 }, "text": "Wild seedling from Cumberland, Ky.", "attrs": { "row": 12, "col": 1, "rowspan": 1, "colspan": 1, "bbox_refit": true } } ] }, { "id": "p2.3", "order": 29, "type": "text", "bbox": { "l": 79.68, "t": 479.52, "r": 338.16, "b": 149.28 }, "text": "isozymes to evaluate the genetic diversity represented in 32 paw-paw cultivars and advanced selections from the breeding program of R. Neal Peterson of the PawPaw Foundation. These authors determined that the isozyme marker variation in cultivated paw-paw is comparable to those of other long-lived temperate woody perennials of widespread geographic range with insect-pollinated outcrossing breeding systems, secondary asexual reproduction and animal-dispersed seed, thus having a higher level of genetic diversity than Rogstad et al. (1991) had reported. Huang et al. (1997) acknowledged that the results may have been impacted by non-random selection because several of the trees studied may have been purposely selected by pawpaw enthusiasts for desirable characteristics such as large fruit size or good growth vigor. Using isozymes, Huang et al. (1998) also assessed the level of genetic diversity within wild collected pawpaw accessions at KSU and examined genetic diversity between pawpaw populations from different geographical locations. Isozymes were used to score 23 loci using 25 to 50 trees from each of nine populations. The level of genetic variation found in KSU repository accessions was similar to that found in cultivated pawpaws (Huang et al., 1997). Using 12 randomly amplified polymorphic DNA (RAPD) primers, Huang et al. (2000) identified 21 Mendelian markers and determined that the level of genetic diversity in six populations in the KSU repository collection was higher than determined for pawpaw by the same authors using isozymes (Huang et al., 1997, 1998). Huang et al. (2003) have also used additional RAPD markers for fingerprinting pawpaw cultivars and reported similar levels of genetic diversity in cultivated pawpaw, in terms of Nei's genetic diversity constant ([p2.3.1]), to that reported for wild pawpaw populations by Huang et al. (2000).", "children": [ { "id": "p2.3.1", "order": 30, "type": "text", "bbox": { "l": 79.68, "t": 479.52, "r": 338.16, "b": 149.28 }, "text": "H_{e}", "attrs": { "inline": true, "text_offset": 1800, "text_span": 8, "bbox_estimated": true } } ] }, { "id": "p2.4", "order": 31, "type": "text", "bbox": { "l": 79.44, "t": 150, "r": 338.16, "b": 93.36 }, "text": "Microsatellites, or simple sequence repeats (SSRs), are abundant throughout eukaryotic genomes (Kijas et al., 1995). The inter-simple sequence repeat PCR (ISSR-PCR) methodology uses a primer composed of a specific microsatellite sequence anchored at either the 3' or 5' end to amplify scorable DNA products (Gupta et al., 1994; Zietkiewicz et al., 1994). ISSR-PCR uses longer primers and has higher annealing temperatures than RAPD markers. It also has a higher degree of polymorphisms and reproducibility than do RAPD markers (Fang and Roose, 1997; Meyer et al., 1993). SSRs have become the marker system of choice for genetic diversity studies of many crops, such as apple [Malus × sylvestris (L.) Mill. Var. domestica (Borkh) Mansf.] (Gianfranceschi et al., 1998) and pear (Pyrus L.) (Yamamoto et al., 2001), due to their abundance in the genome and high degree of polymorphism (Weber and May, 1989). ISSR-PCR, which uses the occurrence of SSRs in the plant genome, has been used successfully to characterize genetic diversity within and among plant populations (Qian et al., 2001). The primary objectives of this study were to identify ISSR markers that segregate in a simple Mendelian fashion and use these markers to evaluate the genetic diversity in commercially available pawpaw cultivars. We were also interested in comparing diversity estimates with those previously reported for pawpaw using isozymes and RAPD markers." }, { "id": "p2.5", "order": 32, "type": "text", "bbox": { "l": 344.64, "t": 776.88, "r": 603.6, "b": 576.72 }, "text": "et al., 1994; Zietkiewicz et al., 1994). ISSR-PCR uses longer primers and has higher annealing temperatures than RAPD markers. It also has a higher degree of polymorphisms and reproducibility than do RAPD markers (Fang and Roose, 1997; Meyer et al., 1993). SSRs have become the marker system of choice for genetic diversity studies of many crops, such as apple [Malus × sylvestris (L.) Mill. Var. domestica (Borkh) Mansf.] (Gianfranceschi et al., 1998) and pear (Pyrus L.) (Yamamoto et al., 2001), due to their abundance in the genome and high degree of polymorphism (Weber and May, 1989). ISSR-PCR, which uses the occurrence of SSRs in the plant genome, has been used successfully to characterize genetic diversity within and among plant populations (Qian et al., 2001). The primary objectives of this study were to identify ISSR markers that segregate in a simple Mendelian fashion and use these markers to evaluate the genetic diversity in commercially available pawpaw cultivars. We were also interested in comparing diversity estimates with those previously reported for pawpaw using isozymes and RAPD markers." }, { "id": "p2.6", "order": 33, "type": "heading", "bbox": { "l": 423.12, "t": 566.88, "r": 526.08, "b": 555.84 }, "text": "Materials and Methods", "attrs": { "block_label": "paragraph_title" }, "level": 3 }, { "id": "p2.7", "order": 34, "type": "text", "bbox": { "l": 344.64, "t": 546, "r": 603.6, "b": 369.6 }, "text": "PLANT MATERIAL. Dormant cuttings were collected in mid-March from pawpaw trees located at the KSU Repository for Asimina species in Frankfort, Ky., placed in beakers of distilled water, and placed under fluorescent room lighting at room temperature (21 °C) to force bud break and leaf growth. Additional leaves were also collected from plants in the field in mid-April. Leaf material was collected from the 34 progeny of controlled crosses (22 from 1-7-1 x 2-54 and 12 from the progeny of the reciprocal cross) and from the individual parents 1-7-1 and 2-54. For the genetic diversity study, leaf samples were collected from 19 commercially available pawpaw cultivars spanning the native range of pawpaw (Table 1). Leaf samples of the pawpaw cultivars Greenriver Belle, Sue, and IXL were obtained from a private nursery (Nolin River Nut Tree Nursery, Nolin River, Ky.). Samples of all other cultivars were obtained from the KSU repository orchards in Frankfort, Ky." }, { "id": "p2.8", "order": 35, "type": "text", "bbox": { "l": 344.88, "t": 368.4, "r": 603.12, "b": 106.56 }, "text": "DNA EXTRACTION. DNA was extracted from the leaves using the Promega Wizard Genomic DNA purification kit (Promega Co., Madison, Wis.). About 50 mg of leaf tissue was ground in a mortar with dry ice, and transferred to a 1.5 mL microfuge tube. Nuclei lysis solution was added (600 μL/tube), vortexed for 1 to 3 s and samples incubated for 15 min at 65 °C. Two microliters RNase A (10 mg·mL⁻¹; Sigma Chemical Co., St. Louis, Mo.) was added to each sample, and the tube incubated for 15 min at 37 °C. Samples were cooled for 1 to 5 min on ice, and 200 μL protein precipitation solution was added. Samples were vortexed for 20 s, centrifuged (15,800 gₙ) for 5 min, then 750 μL of supernatant was transferred to a tube containing 750 μL 99% isopropanol. Samples were mixed by inversion and centrifuged for 5 min. Supernatants were discarded and the DNA pellets were first washed with 500 μL of 70% ethanol, next with 500 μL of 95% ethanol and finally air dried for 15 min. The DNA was solubilized in 150 μL of 1× TE (10 mM Tris-HCl, 1 mM EDTA, pH 8.0) buffer by an initial incubation at 65 °C for 1 h, then overnight at 4 °C. The samples were centrifuged for 3 min (15,800 gₙ) to remove insoluble material and the supernatants were transferred to new tubes. The DNA concentration and a 260/280 nm absorbance ratio were determined using a GeneQuant RNA/DNA calculator spectrophotometer (Pharmacia Biotech, Cambridge, U.K.). All samples were stored at −80 °C until needed." }, { "id": "p2.9", "order": 36, "type": "text", "bbox": { "l": 357.36, "t": 105.12, "r": 603.6, "b": 94.32 }, "text": "ISSR-PCR AMPLIFICATION. The PCR reaction mixture contained" }, { "id": "p2.10", "order": 37, "type": "footer", "bbox": { "l": 78.48, "t": 73.2, "r": 95.76, "b": 62.4 }, "text": "522", "attrs": { "block_label": "number" } }, { "id": "p2.11", "order": 38, "type": "footer", "bbox": { "l": 426.72, "t": 72.72, "r": 601.68, "b": 62.64 }, "text": "J. AMER. SOC. HORT. SCI. 128(4):521–525. 2003.", "attrs": { "block_label": "footer" } } ], "excluded_furniture": { "first": [], "last": [] } }