Title
アスパラガス忌地現象における生物・化学的因子の相互作
用並びに植物生育改善法の実証的研究( 本文(Fulltext) )
Author(s)
LIU, JIA
Report No.(Doctoral
Degree)
博士(農学) 甲第662号
Issue Date
2017-03-13
Type
博士論文
Version
ETD
URL
http://hdl.handle.net/20.500.12099/56210
※この資料の著作権は、各資料の著者・学協会・出版社等に帰属します。ࢫࣃࣛ࢞ࢫᚷᆅ⌧㇟࠾ࡅࡿ⏕≀࣭ᏛⓗᅉᏊࡢ┦స⏝୪ࡧ
᳜≀⏕⫱ᨵၿἲࡢᐇドⓗ◊✲
2 0 1 6 ᖺ
ᒱ㜧ᏛᏛ㝔㐃ྜ㎰Ꮫ◊✲⛉
⏕≀⏕⏘⛉Ꮫ
㸦ᒱ 㜧 Ꮫ㸧
ี
֣
ࢫࣃࣛ࢞ࢫᚷᆅ⌧㇟࠾ࡅࡿ⏕≀࣭ᏛⓗᅉᏊࡢ┦స⏝୪ࡧ
᳜≀⏕⫱ᨵၿἲࡢᐇドⓗ◊✲
┠ ḟ ➨1 ❶ ᚷᆅ⌧㇟࠾ࡅࡿᏛⓗ࣭⏕≀ⓗᅉᏊࡢ┦స⏝ゎᯒ ࣭࣭࣭㸯 ➨1 ⠇ ࣞࣟࢣ࣑࢝ࣝ❧ᯤ⳦ࡢ┦స⏝㛵ࡍࡿ in vitro ホ౯ ➨2 ⠇ ࣞࣟࢣ࣑࢝ࣝࡀࢫࣃࣛ࢞ࢫ࠾ࡅࡿ❧ᯤⓎㄏᑟཬࡰࡍ ᙳ㡪ࡢin vitro ホ౯ ➨2 ❶ ࢫࣃࣛ࢞ࢫ❧ᯤ࠾ࡅࡿ⪏ᛶㄏᑟ ࣭࣭࣭15 ➨1 ⠇ ࢫࣃࣛ࢞ࢫ࡛ࡢ AMF ࡼࡿ❧ᯤ⪏ᛶㄏᑟࡢရ✀㛫ᕪ
➨2 ⠇ Split root system ἲࡼࡿࢫࣃࣛ࢞ࢫ࡛ࡢ⏕≀࣭Ꮫⓗᡭἲࡼࡿ ❧ᯤㄏᑟᢠᛶホ౯ཬࡧᢠ㓟ᶵ⬟ኚືゎᯒ ➨3 ❶ ࢫࣃࣛ࢞ࢫᨵ᳜㞀ᐖᅡ࡛ࡢ⏕≀࣭Ꮫⓗᡭἲࡼࡿ⥲ྜⓗ᳜≀⏕⫱ᨵၿࡢ᳨ド࣭࣭࣭77 ➨1 ⠇ ᨵ᳜ 12 㐌㛫ᚋ࠾ࡅࡿⓎ࣭⏕⫱ᨵၿホ౯ ➨2 ⠇ ᨵ᳜ᚋ࠾ࡅࡿ㔞ཬࡧ✭ⱼࡢᢠ㓟≀㉁ྵ㔞ホ౯ せ ࣭࣭࣭102 ㅰ ㎡ ࣭࣭࣭105 ᘬ⏝ᩥ⊩ ࣭࣭࣭106
1
➨
1 ❶
ᚷᆅ⌧㇟࠾ࡅࡿᏛⓗ࣭⏕≀ⓗᅉᏊࡢ┦స⏝ゎᯒ
2 ⥴ ゝ
ᚷᆅ⌧㇟㸦㐃స㞀ᐖ㸧ࡣᅬⱁస≀⏕⏘࠾࠸࡚ࠊ᱂ᇵ୰ᚋᮇཬࡧᨵ᳜ᚋ⏕⫱Ⰻࠊ㔞࣭ရ
㉁పୗࡀⓎ⏕ࡍࡿ⌧㇟࡛ࠊ㔝⳯࠾ࡅࡿࡋ࡚ࠊࢫࣃࣛ࢞ࢫ࡛ࡣᅜෆእࡢ⏘ᆅ࡛῝้ࡋࠊ
⏘ᆅࡁ࡞⿕ᐖࢆ࠼࡚࠸ࡿ㸦Wong and Jeffries㸪2006㸹Knaflewski ࡽ㸪2008; Elmer, 2015㸧ࠋ
ᚷᆅ⌧㇟ࡢⓎ⏕ࡣ⏕≀ⓗᅉᏊ࡛࠶ࡿᅵተᐖ㸦Wong and Jeffries㸪2006㸹Knaflewski ࡽ㸪2008㸧
ࡸ㠀⏕≀ⓗᅉᏊࡢࣞࣟࣃࢩ࣮ࠊ㣴ศᆒ⾮➼㸦Yong㸪1984㸹Miller ࡽ㸪1991㸹Lake ࡽ㸪1993㸹
᪥➟㸪2000㸧ࡀ㛵ࢃࡗ࡚࠸ࡿ⪃࠼ࡽࢀ࡚࠸ࡿࠋせ࡞⏕≀ⓗᅉᏊࡋ࡚ࡣࣇࢨ࣒ࣜ࢘⳦ࡼࡿ
ᐖࡀุ᫂ࡋ࡚࠾ࡾࠊFusarium oxysporum f. sp. asparagi㸦Foa㸧ࡸ F. proliferatum㸦Fp㸧ࠊF. redolens
➼ࡀᣦࡉࢀ࡚࠸ࡿ㸦Reid ࡽ㸪2001㸹Wong and Jeffries㸪2006㸹Knaflewski ࡽ㸪2008㸧ࠋ୍᪉ࠊ᪥
ᮏᅜෆࡢᚷᆅᅡሙ࠾ࡅࡿࣇࢨ࣒ࣜ࢘⳦✀ࡘ࠸࡚ࡣ Foa ཬࡧ Fp ࡀඃ༨✀࡛࠶ࡾࠊࡑࢀࡽࡼ ࡿ❧ᯤ࠾ࡼࡧᰴ⭉ࡀせ࡞ࣇࢨ࣒ࣜ࢘ᐖ⪃࠼ࡽࢀ࡚࠸ࡿ㸦Nahiyan ࡽ㸪2011㸧ࠋ ᏛⓗᅉᏊࡋ࡚ࡣࣞࣟࣃࢩ࣮ࡀᣦࡉࢀࠊࢫࣃࣛ࢞ࢫ࡛ࡣ᰿ࡽศἪࡉࢀࡿᏛ≀㉁㸦 ࣞࣟࢣ࣑࢝ࣝ㸧ࡀ⮬㌟ࡔࡅ࡛࡞ࡃᨵ᳜ᚋࡢಶయࡢ⏕⫱ࢆ㜼ᐖࡍࡿࡇࡀሗ࿌ࡉࢀ࡚࠸ࡿ㸦Lake ࡽ㸪 1993㸹ඖᮌࡽ㸪2006㸧ࠋࡑࢀࡽࣞࣟࢣ࣑࢝ࣝࡣỈ⁐ᛶࠊ⇕Ᏻᐃᛶ࡛ࠊ᱂ᇵ⤊ᚋᅡሙṧᏑࡍ ࡿ᰿ṧ´ࡽࡶⁱฟࡋࠊᅡሙ✚ࡋ࡚ࣞࣟࣃࢩ࣮ࢆⓎ⏕ࡉࡏ࡚࠸ࡿ⪃࠼ࡽࢀ࡚࠸ࡿ㸦Yang㸪 1982㸹Hartung and Stephens㸪1983㸧ࠋࣞࣟࢣ࣑࢝ࣝ⪃࠼ࡽࢀ࡚࠸ࡿᏛ≀㉁ࡣከࡃࠊࡇࢀࡲ࡛
ࠊ࢝ࣇ࢙㓟ࠊࣇ࢙ࣝࣝ㓟ࠊࢣࣝࢭࢳࣥࠊࣜࣥࢦ㓟ࠊࢡ࢚ࣥ㓟➼ࡀ♧၀ࡉࢀ࡚࠸ࡿ㸦Hartung ࡽ㸪
1990㸹Miller ࡽ㸪1991㸹Lake ࡽ㸪1993㸹 Inderjit ࡽ 1995㸧ࠋ୍᪉ࠊᚷᆅ⌧㇟ࡢせᅉࡋ࡚๓㏙ ࡢ⏕≀ⓗ࣭ᏛⓗᅉᏊࡢᏑᅾࡀᣲࡆࡽࢀ࡛࠸ࡿࡀࠊࡑࢀࡽ༢⊂ᅉᏊࡘ࠸࡚ࡢ᳨ウࡀከࡃࠊ ⏕≀ⓗ࣭ᏛⓗᅉᏊ㛫ࡢ┦స⏝ࡘ࠸࡚ࡣ᫂࡞Ⅼࡀከ࠸ࠋࡼࡗ࡚ࠊࢫࣃࣛ࢞ࢫᚷᆅ⌧㇟ ࠾ࡅࡿࣞࣟࣃࢩ࣮ᐖࡢ┦స⏝㛵ࡍࡿ᳨ウࡣࠊࢫࣃࣛ࢞ࢫᚷᆅ⌧㇟ࡢⓎ⏕ᶵᵓゎ᫂ཬ ࡧ㜵㝖ᑐ⟇࠾࠸࡚ࡶᚲせࡘ㔜せ࡛࠶ࡿ⪃࠼ࡽࢀࡿࠋ ᮏ◊✲࡛ࡣࠊࢫࣃࣛ࢞ࢫᚷᆅ⌧㇟Ⓨ⏕ᶵᵓ࠾ࡅࡿ⏕≀ⓗᅉᏊᏛⓗᅉᏊࡢ┦స⏝ࡘ ࠸࡚ࡢ▱ぢࢆᚓࡿࡇࢆ┠ⓗࡋࠊࢫࣃࣛ࢞ࢫ❧ᯤ4 ✀ࡢࣞࣟࢣ࣑࢝ࣝ♧၀≀㉁ࡢ┦ స⏝ࡘ࠸࡚in vitro ホ౯ࢆ⾜ࡗࡓࠋ
3 ᮦᩱཬࡧ᪉ἲ
➨1 ⠇ ࣞࣟࢣ࣑࢝ࣝ❧ᯤ⳦ࡢ┦స⏝㛵ࡍࡿ in vitro ホ౯
Potato dextrose agar㸦PDA㸧ᇵᆅ࡛⣧⢋ᇵ㣴ࡋࡓࢫࣃࣛ࢞ࢫ❧ᯤ⳦㹙Fusarium oxysporum f. sp.
asparagi㸦Foa : MAFF 305556㸧㹛ࡢ⳦ࡑ࠺ࢆ Czapec-Dox㸦➨ 1 ⾲, Czapek, 1902-1903; Dox, 1910㸧
ᾮయᇵᆅΰྜࡋࠊேᕤẼ㇟ჾෆ㸦25Υᬯᡤ㸧࡛ᇵ㣴ࡋࠊFoa ศ⏕⬊Ꮚᠱ⃮ᾮࡋࡓࠋ⥆࠸࡚ࠊ ᪂ࡓ࡞ Czapek-Dox ᾮయᇵᆅࢆసᡂࡋࠊᩘ✀ࣞࣟࢣ࣑࢝ࣝ♧၀≀㉁㸦࢝ࣇ࢙㓟㸪ࣇ࢙ࣝࣝ㓟㸪 ࢣࣝࢭࢳࣥ㸪ࣜࣥࢦ㓟㸧ࢆࢡ࣮ࣜࣥ࣋ࣥࢳෆ࡛ῧຍ㸦0.01 㸪0.1%㸪w/v㸧ࡋࡓࠋᑐ↷༊ࡋ࡚ ␃Ỉῧຍ༊ࢆタࡅࡓࠋྛࣞࣟࢣ࣑࢝ࣝࢆῧຍࡋࡓCzapec-Dox ᾮయᇵᆅ Foa ศ⏕⬊Ꮚᠱ⃮ᾮࢆ ຍ࠼ࠊேᕤẼ㇟ჾෆ㸦25Υ㸪ᬯᡤ㸧࡛ 1 㐌㛫࠺ᇵ㣴㸦100rpm㸧ࡋࡓࠋྛฎ⌮༊ࡀᩘࡣ 5 ࡋࠊᇵ㣴⤊ᚋࠊ⾑⌫ィ⟬┙ࢆ⏝࠸࡚ศ⏕⬊Ꮚᩘࢆィ ࡋࠊᑐ↷༊ᑐࡍࡿྛࣞࣟࢣ࣑࢝ࣝ ῧຍ༊ࡢቑṪᣦᩘࢆ⟬ฟࡋࡓࠋ ➨2 ⠇ ࣞࣟࢣ࣑࢝ࣝࡀࢫࣃࣛ࢞ࢫ࠾ࡅࡿ❧ᯤⓎㄏᑟཬࡰࡍᙳ㡪ࡢ in vitro ホ౯
Knop’s ᇵᆅ㸦Blok and Bollen, 1996, ➨ 2 ⾲㸧➨ 1 ⠇ྠᵝࡢࣞࣟࢣ࣑࢝ࣝ♧၀≀㉁ ࢆῧຍ㸦0.01㸪0.1%㸪w/v㸧ࡋࠊࣞࣟࢣ࣑࢝ࣝῧຍ Knop’s ᇵᆅࢆసᡂࡋࡓࠋࢫࣃࣛ࢞ࢫ㸦Asparagus officinalis L., ‘Welcome’㸧ࢆࢡ࣮ࣜࣥ࣋ࣥࢳෆ࡛ 70㸣࢚ࢱࣀ࣮ࣝ 10 ⛊ࠊࣥࢳ࣑࣍ࣝࣥ㸦᭷ຠ ሷ⣲1%㸧 15 ศᾐₕࡋ࡚⾲㠃ẅ⳦ࡋࠊ␃Ỉ࡛Ὑίᚋࠊࣞࣟࢣ࣑࢝ࣝῧຍ Knop’s ᇵᆅ ✀ࡋࡓࠋྛヨ㦂༊20 ಶయࡢ 2 ࡋࠊᑐ↷༊ࡣࣞࣟࢣ࣑࢝ࣝ↓ῧຍࡢ Knop’s ᇵᆅࢆ⏝ ࡋࡓࠋᇵ㣴ࡣேᕤẼ㇟ჾ㸦LPH-241SP, ᪥ᮏ་ᶵᲔ〇సᡤ㸧ෆ࡛⾜࠸ࠊⓎⱆࡲ࡛ࡣ 25Υ࣭ᬯᡤࠊ Ⓨⱆᚋࡣ25Υ࣭16 㛫᪥㛗ࡋࡓࠋ✀ 4 㐌㛫ᚋ⏕⫱ㄪᰝࢆ⾜ࡗࡓࠋ⥆࠸࡚ࠊFoa ࢆቑṪࡉࡏ ࡓPDA ᐮኳ∦㸦5×5mm㸧ࢆᐇ⏕ᇶ㒊᥋✀ࡋࡓ㸦➨ 1 ᅗ㸧ࠋFoa ᥋✀ 2 㐌㛫ᚋࠊྛಶయࡢ㈓ⶶ᰿
࠾ࡅࡿ⨯㛗ࢆィ ࡋࠊ㛗ᑐࡍࡿ⨯㛗ᣦᩘ㸦IDL㸸Index of diseased length in total storage root
length㸧ࢆ௨ୗࡢᘧࡼࡾ⟬ฟࡋࡓࠋ
4 ྛಶయࡢ㈓ⶶ᰿㛗ᑐࡍࡿ⨯㛗ࡢྜ 0㸸0% 1㸸~20% 2㸸20~40% 3㸸40~60% 4㸸60~80% 5㸸80~100% ౪ヨಶయᩘ× 5 (᭱ᣦᩘ) ౪ヨಶయᩘ × 5 (᭱ᣦᩘ) IDL㸻× 100DL㸻 ∑ಶయᩘ× ᣦᩘ
NaNO΄
KHPO΅
MgSO΅䞉7HO
KCl
FeSO΅䞉7HO
Sucrose 㻌
Table 1. Composition of Czapek-Dox medium.
pH 㻌 㻌 㻌 5.8
3g/l
1g/l
0.5g/l
0.5g/l
0.01g/l
30g/l
5KNO΄
Ca(NO΄)䞉7HO
KHPO΅
MgSO΅䞉7HO
FeSO΅䞉2HO
Sucrose㻌 㻌
Agar㻌 㻌 㻌 㻌 㻌 㻌
Table 2. Composition of Knop’s medium.
0.2g/l
1.15g/l
0.2g/l
0.2g/l
0.12g/l
30g/l
12.5g/l
6Fig.1㻌 Asparagus seedlings inoculated with Fusarium oxysporum f. sp. asparagi (Foa).㻌
Foa
8 ⤖ ᯝ ➨1 ⠇ ࣞࣟࢣ࣑࢝ࣝ❧ᯤ⳦ࡢ┦స⏝㛵ࡍࡿ in vitro ホ౯ ࡚ࡢࣞࣟࢣ࣑࢝ࣝ♧၀≀㉁ῧຍ༊࠾࠸࡚ࠊᑐ↷༊ᑐࡍࡿ❧ᯤ⳦ቑṪᣦᩘࡣ100 ࢆୗ ᅇࡗ࡚࠸ࡓࠋ㸦➨2 ᅗ㸧ࠋFoa ቑṪᣦᩘࡣࣞࣟࢣ࣑࢝ࣝࡢ✀㢮ཬࡧ⃰ᗘࡼࡗ࡚ኚືࡋࡓࠋࡇࡢ ሙྜࠊ4 ✀ࣞࣟࢣ࣑࢝ࣝῧຍ༊࠾࠸࡚ࠊቑṪᣦᩘࡀ᭱ࡶ㧗ࡗࡓࡢࡣࣜࣥࢦ㓟ཬࡧࣇ࢙ࣝࣝ 㓟ࡢ0.01%ῧຍ༊࡛࠶ࡾࠊ᭱ࡶపࡗࡓࡢࡣ࢝ࣇ࢙㓟ཬࡧࢣࣝࢭࢳࣥࡢ 0.1%ῧຍ༊࡛࠶ࡗࡓࠋ ➨2 ⠇ࣞࣟࢣ࣑࢝ࣝࡀࢫࣃࣛ࢞ࢫ࠾ࡅࡿ❧ᯤⓎㄏᑟཬࡰࡍᙳ㡪ࡢ in vitro ホ౯ ✀4 㐌㛫ᚋ⏕⫱ㄪᰝࡢ⤖ᯝࠊᆅୖ㒊≀㔜ࡣࢇࡢࣞࣟࢣ࣑࢝ࣝῧຍ༊࡛ࡁࡃῶᑡ ࡋ࡚࠸ࡓࡀࠊࣜࣥࢦ㓟0.01㸣ῧຍ༊࡛ࡣቑຍࡋ࡚࠸ࡓ ➨ 3 ᅗ ࠋᆅୗ㒊≀㔜ࡣࠊ࢝ࣇ࢙㓟ࠊࢣ ࣝࢭࢳࣥ0.1%ῧຍ༊ཬࡧ࢝ࣇ࢙㓟ࠊࣇ࢙ࣝࣝ㓟 0.01%ῧຍ༊࡛ῶᑡࡋ࡚࠸ࡓࡀࠊࣇ࢙ࣝࣝ㓟 0.1% ῧຍ༊࡛ࡣቑຍࡋ࡚࠸ࡓࠋࡢࣞࣟࢣ࣑࢝ࣝῧຍ༊࠾࠸࡚ᑐ↷༊ࡢᕪࡣࡳࡽࢀ࡞ࡗࡓࠋ ❧ᯤ⳦᥋✀2 㐌㛫ᚋࠊⓎ⋡ࡘ࠸࡚ࡣࠊᑐ↷༊ࡼࡾࣞࣟࢣ࣑࢝ࣝῧຍ༊࡛ቑࡍࡿሙྜࡀ ከࡗࡓ ➨ ᅗ ࠋIDL ࡣከࡃࡢࣞࣟࢣ࣑࢝ࣝῧຍ༊࡛ᑐ↷༊ࡼࡾࡶ㧗࠸್ࢆ♧ࡋࡓ㸦➨ 5 ᅗ㸧ࠋ ≉ࠊ࢝ࣇ࢙㓟࣭ࣇ࢙ࣝࣝ㓟0.01%ῧຍ༊࡛ࡣࠊIDL ࡀ 60 ࢆ㉸࠼࡚࠸ࡓࠋ
0
20
40
60
80
100
Index of
Foa
propagat
io
n
a
ab
e
b
c
de
b
d
Fig. 2. Influence of allechemicals on propagation of Foa (Fusarium oxysporum f.
sp. asparagi; MAFF305556). , 0.01%; ᴾ ᴾ , 0.1%. Bars represent standard
errors (n=3). Columns denoted by different letters indicate significant difference
according to Tukey's test (P<0.05).
Caffeic acid Ferulic acid Quercetin Malic acid 9
0
5
10
15
20
25
0
5
10
15
20
D
ry
w
eight
of
shoots
(m
g)
D
ry
w
eight
of
roots
(m
g)
Fig. 3.㻌 Dry weight of asparagus seedlings 4 weeks after sowing in
allelochemical-added Knop’s media. , control; , 0.1%; , 0.01%. Bars represent standard
errors (n=5). Columns denoted by different letters indicate significant difference
according to Tukey's test (P<0.05).
b
c
d
d
cd
bc
c
b
a
b
c c
c
ab
c
bc
a
bc
control Caffeic acid Ferulic acid Quercetin Malic acid 10Incidence of
F
usarium
root
rot
(%)
Fig. 4. Incidence of Fusarium root rot in asparagus plants 2 weeks after Fusarium
oxysporum f. sp. asparagi (MAFF305556) inoculation. Ratio of diseased storage roots
in a root system; 㻌 㻌 , -20; , 20-40; , 40-60; , 60-80; , 80-100 (%).
0
20
40
60
80
100
0.01% 0.1% 0.01% 0.1% 0.01% 0.1% 0.01% 0.1% control Caffeicacid Ferulic acid Quercetin
Malic acid
0
20
40
60
80
IDL
Fig. 5. Index of diseased length to total root length (IDL) in asparagus plants
cultured by allelochemical-added media. , control; , 0.1%; , 0.01%.
Bars represent standard errors (n=10). Columns denoted by different letters
indicate significant difference according to Tukey's test (P<0.05).
a
a
b
c
c
e
d
d
d
control Caffeic acid Ferulic acid Quercetin Malic acid 1213 ⪃ ᐹ ᮏ◊✲࡛ࡣࠊࢫࣃࣛ࢞ࢫᚷᆅ⌧㇟Ⓨ⏕ᶵᵓゎ᫂ࡢ୍⎔ࡋ࡚ࠊ⏕≀ⓗ࣭ᏛⓗᅉᏊࡢ┦స ⏝ࡢin vitro ホ౯ࢆ⾜ࡗࡓࠋ➨ ⠇࡛ࡣࠊCzapek-Dox ᾮయᇵᆅࢆ⏝࠸࡚ࠊ4 ✀㢮ࡢࣞࣟࢣ࣑࢝ࣝ ♧၀≀㉁ࡀࢫࣃࣛ࢞ࢫ❧ᯤ⳦ࡢቑṪཬࡰࡍᙳ㡪ࢆㄪᰝࡋࡓࠋࡑࡢ⤖ᯝࠊᅇ⏝ࡋࡓࣞ ࣟࢣ࣑࢝ࣝ♧၀≀㉁ࡣᮏᐇ㦂⏝࠸ࡓ❧ᯤ⳦ࡢቑṪࢆಁ㐍ࡉࡏࡿࡶࡢࡣ࡞ࡃࠊ࡚ᢚไഴྥ ࡀ࠶ࡗࡓࠋHong-sheng ࡽ㸦2010㸧ࡣࣞࣟࢣ࣑࡛࢝ࣝ࠶ࡿࣇ࢙ࣝࣝ㓟ࡢῧຍࡼࡾ Fusarium
oxysporum f. sp. niveum ࡢ in vitro ࡛ࡢቑṪࡀᢚ࠼ࡽࢀࠊᢚไ⛬ᗘࡣࣇ࢙ࣝࣝ㓟⃰ᗘࡼࡗ࡚␗࡞
ࡾࠊ㧗⃰ᗘ࡛ᙉࡃ࡞ࡿࡇࢆሗ࿌ࡋ࡚࠸ࡿࠋᮏᐇ㦂࠾࠸࡚ࡶྠᵝࡢ⤖ᯝࡀᚓࡽࢀࠊᅇ⏝ࡋ ࡓ4 ✀ࣞࣟࢣ࣑࢝ࣝ࠾࠸࡚ࡣࠊ❧ᯤ⳦ࡢቑṪࢆᢚไࡍࡿ⪃࠼ࡽࢀࡿࠋࡇࡢሙྜࠊῧຍࡍ ࡿࣞࣟࢣ࣑࢝ࣝࡢ✀㢮ཬࡧ⃰ᗘࡼࡾࠊᢚไ⛬ᗘᕪࡀ࠶ࡿࡇࡀ♧၀ࡉࢀࡓࠋ ➨2 ⠇࡛ࡣࠊࣞࣟࢣ࣑࢝ࣝ♧၀≀㉁ࡀࢫࣃࣛ࢞ࢫ❧ᯤⓎཬࡰࡍᙳ㡪ࢆ in vitro ࡛ㄪᰝ ࡋࡓࠋ❧ᯤ⳦᥋✀๓࠾࠸࡚ࠊࣞࣟࢣ࣑࢝ࣝῧຍ༊ࡢ୰ࡣ≀㔜ࡀపୗࡋࠊⱼࡸ᰿ࡢ⏕⫱ ࡀⴭࡋࡃ㜼ᐖࡉࢀ࡚࠸ࡓࡶࡢࡶࡳࡽࢀࡓࠋ≉ࠊ࢝ࣇ࢙㓟࡛ࡣῧຍ⃰ᗘࡶ㛵ࢃࡽࡎࠊⱼࡸ᰿ࡢ ⏕⫱ࡀ᭱ࡶᢚไࡉࢀ࡚࠸ࡓࠋࡋࡋࠊࣜࣥࢦ㓟0.01㸣ῧຍ༊ࡢᆅୖ㒊ཬࡧᆅୗ㒊ࡢ≀㔜ࡣᑐ↷ ༊ࡼࡾቑຍࡋ࡚࠸ࡓࠋࡑࡢࡓࡵࠊ➨1 ⠇ྠᵝࣞࣟࢣ࣑࢝ࣝࡼࡿ᳜≀యࡢ⏕⫱㜼ᐖࡣ⃰ ᗘࡼࡾࡶࣞࣟࢣ࣑࢝ࣝࡢ✀㢮ࡢᙳ㡪ࡀࡁ࠸ࡇࡀ♧၀ࡉࢀࡓࠋࣞࣟࢣ࣑࢝ࣝࡢࢫࣃࣛ࢞ ࢫᑐࡍࡿⓎⱆᢚไࡣࠊMiller ࡽ㸦1991㸧ࡀሗ࿌ࡋ࡚࠸ࡿࠋࡲࡓࠊࡢࣞࣟࢣ࣑࡛࢝ࣝ࠶ࡿࠊ ࢩࢼ࣑ࣥ㓟ࠊࢩࣗ࢘㓟ࠊࣇ࢙ࣝࣝ㓟࡞ࡣࣞࢱࢫ࡞ࡢࡢ᳜≀࠾࠸࡚Ⓨⱆࡀ㜼ᐖࡉࢀࡿࡇ ࡀ☜ㄆࡉࢀ࡚࠾ࡾ㸦⊦㇂ࡽ1999㸧ࠊࣞࣟࢣ࣑࢝ࣝࡢ✀㢮ࡼࡗ࡚ᢚไ⛬ᗘࡀ␗࡞ࡿࡇ࡞ࡶ ♧၀ࡉࢀ࡚࠸ࡿ㸦Cheaves ࡽ㸪2003㸧ࠋ ᮏᐇ㦂࠾ࡅࡿIDL ࡼࡿⓎホ౯࡛ࡣࠊከࡃࡢࣞࣟࢣ࣑࢝ࣝῧຍ༊࡛Ⓨࡀ㔜ᗘ࡞ࡿഴ ྥࡀࡳࡽࢀࡓࠋࡼࡗ࡚ࠊ➨1 ⠇ࡢ⤖ᯝࡶྵࡵࠊࣞࣟࢣ࣑࢝ࣝࡀ᳜≀యࢆࡋ࡚Ⓨࢆຓ㛗ࡍࡿ ྍ⬟ᛶࡀ♧၀ࡉࢀࡓࠋᅇࠊࣞࣟࢣ࣑࢝ࣝ 0.1%ῧຍ༊࠾࠸࡚ࡣࠊ᰿ఙ㛗ࡀᢚไࡉࢀ࡚࠸ࡓࠋ ࡑࡢࡓࡵࠊ᰿ࡢᡂ㛗ࡀ༑ศ࡞≧ែ࡛❧ᯤឤᰁࡋࡓࡓࡵࠊⓎࡀ㔜ᗘ࡞ࡗࡓྍ⬟ᛶࡀ࠶ࡿࠋ ࡋࡋࠊᑐ↷༊᰿㛗ࡁ࡞ᕪࡀࡳࡽࢀ࡞ࡗࡓࣞࣟࢣ࣑࢝ࣝࡢ 0.01%༊࠾࠸࡚ࡶⓎᗘ
14 ࡀ㧗ࡃ࡞ࡗ࡚࠸ࡓࠋࡇࡢࡇࡽࠊ᰿ఙ㛗௨እࡢせ⣲ࡶࣞࣟࢣ࣑࢝ࣝࡀᙳ㡪ࡋ࡚࠸ࡿࡇࡀ ⪃࠼ࡽࢀࡓࠋSang ࡽ㸦2002㸧ࡣࠊࣞࣟࢣ࣑࢝ࣝᏑᅾୗ࡛ࡢ᰿㛗ࡔࡅ࡛࡞ࡃࠊ᰿➃ࡢᙧែ╔┠ ࡋࠊࣞࣟࢣ࣑࢝ࣝᏑᅾୗ࡛ࡢ᰿➃ࡢ┤ᚄࡀࡁࡃ࡞ࡿࡇࡸࠊ⓶ᒙࡀ⭾ࡽࢇ࡛࠸ࡓࡇࢆሗ࿌ ࡋ࡚࠸ࡿࠋࡲࡓࠊ࢝ࣇ࢙㓟ࢆῧຍࡍࡿࡇ࡛ࢲࢬ㸦Glycine max㸧ࡢ᰿ࡢఙ㛗ᢚไࡶ㓝⣲ άᛶࡀኚࡋࠊᮌࡀ㐍ࡴ࠸࠺ሗ࿌ࡶ࠶ࡿ㸦Gisele ࡽ.㸪2011 㸧ࠋࡇࢀࡽࡢࡇࡽࠊ⤌⧊ᵓ㐀 ࡸ௦ㅰࡶࣞࣟࢣ࣑࢝ࣝࡀఱࡽࡢస⏝ࢆཬࡰࡋ࡚࠸ࡿࡢ࡛ࡣ࡞࠸⪃࠼ࡽࢀࠊ᰿ᙧែࡸ᰿ ෆᡂศࡘ࠸࡚ࡶ╔┠ࡋㄪᰝࡍࡿᚲせࡀ࠶ࡿ⪃࠼ࡽࢀࡓࠋ ➨1 ⠇ཬࡧ➨ 2 ⠇࡛⏝ࡋࡓᩘ✀ࣞࣟࢣ࣑࢝ࣝ♧၀≀㉁ࡣࠊࢫࣃࣛ࢞ࢫࡢ㈓ⶶ᰿ࡲࡓࡣ᰿ ᢳฟᾮྵࡲࢀ࡚࠸ࡿ≀㉁࡛ࠊࡲࡓࠊࡢ᳜≀࠾࠸࡚ࣞࣟࣃࢩ࣮≀㉁࡛࠶ࡿࡇࡶ♧၀ࡉࢀ ࡚࠾ࡾࠊ⥲ྜⓗࢫࣃࣛ࢞ࢫࡢࣞࣟࣃࢩ࣮♧၀≀㉁⪃࠼ࡽࢀ࡚࠸ࡿࡶࡢࢆ㑅ᢤࡋ࡚⏝ࡋ
ࡓࠋ㸦Hartung ࡽ㸪1990㸹Miller ࡽ㸪1991㸹 Inderjit㸪1995㸹⊦㇂ࡽ㸪1999㸹Hong-sheng ࡽ㸪2009㸧ࠋ
ࡋࡋࠊᅇ⏝ࡋࡓࡢࡣࠊࣇ࢙ࣀ࣮ࣝ㓟ࡸ᭷ᶵ㓟࡛ࠊ࣑ࣀ㓟ࡣ⏝ࡋ࡚࠸࡞࠸ࠋ࣑ࣀ㓟࡞
࡛ࡣࠊFusarium ࡢ⬊Ꮚ㔞ࡀቑຍࡋࡓ࠸࠺ሗ࿌ࡶ࠶ࡿ㸦Hong-Sheng㸪2010㸧ࠋࡲࡓࠊHong-Sheng
ࡽ㸦2012㸧ࡣࠊࢫ࢝㸦Citrullus lanatus㸧࠾࠸࡚ Fusarium ᐖࡢឤཷᛶရ✀ከࡃྵࡲࢀ࡚࠸
ࡓࣂࢽࣜࣥ㓟㸪ࢩࣜࣥ࢞㓟࡛ࡣࠊFusarium ࡢ⳦⣒ࡀࡼࡃⓎ㐩ࡋࡓࡶሗ࿌ࡋ࡚࠾ࡾࠊࡢࣞࣟ ࢣ࣑࢝ࣝ࡞࡛❧ᯤ⳦ࡀቑṪࡍࡿྍ⬟ᛶࡣ༑ศ⪃࠼ࡽࢀࡿࠋࡲࡓࠊᅇࡣࣞࣟࢣ࣑࢝ࣝࡀ ❧ᯤཬࡰࡍᙳ㡪࡛࠶ࡾࠊ❧ᯤⓎࡀࣞࣟࢣ࣑࢝ࣝཬࡰࡍᙳ㡪ࡣㄪᰝࡋ࡚࠸࡞࠸ࠋBlok ࡽ㸦1997㸧❧ᯤ⳦ឤᰁࡼࡾࣞࣟࣃࢩ࣮ࡀⓎ⏕ࡍࡿࡇࢆ♧၀ࡋ࡚࠸ࡿࠋࡼࡗ࡚ࠊࣞࣟࢣ ࣑࢝ࣝ࠾࠸࡚ࡶཎ⳦ࡢឤᰁࡼࡿኚࡀ࠶ࡿྍ⬟ᛶࡶ⪃࠼ࡽࢀࡿࠋ ௨ୖࡢࡇࡽࠊࣇ࢙ࣀ࣮ࣝ㓟ࡸ᭷ᶵ㓟࡞ࡢࣞࣟࢣ࣑࢝ࣝ♧၀≀㉁࡛ࡣࠊࢫࣃࣛ࢞ࢫ❧ ᯤ⳦ࡢቑṪࢆಁࡍࡢ࡛ࡣ࡞ࡃࠊ᳜≀యࢆࡋࡓ㛫᥋ⓗ࡞⤒㊰࡛Ⓨస⏝ࡍࡿ⪃࠼ࡽࢀࡓࠋ ࡲࡓࠊᚋࡣFusarium ࡞ࡢཎ⳦ࡀࣞࣟࢣ࣑࢝ࣝศἪ㔞࡞ཬࡰࡍᙳ㡪࡞ࡶㄪᰝࡍࡿᚲ せࡀ࠶ࡿ⪃࠼ࡽࢀࡿࠋ
15
➨
2 ❶
ࢫࣃࣛ࢞ࢫ❧ᯤ࠾ࡅࡿ⪏ᛶㄏᑟ
16 ⥴ ゝ ᚷᆅ⌧㇟ࡣᅬⱁస≀⏕⏘࠾࠸࡚ࠊ⤒῭᱂ᇵ୰ᚋᮇཬࡧᨵ᳜ᚋ⏕⫱Ⰻࠊ㔞࣭ရ㉁పୗࡀ Ⓨ⏕ࡍࡿ⌧㇟࡛ࠊ㐃స㞀ᐖࡢࡼ࠺≉ከᖺ⏕స≀࡛Ⓨ⏕ࡋ࡚࠸ࡿࠋ㔝⳯࠾ࡅࡿࡋ࡚ࠊ ࢫࣃࣛ࢞ࢫ࡛ࡣᚷᆅ⌧㇟ࡀᅜෆእࡢ⏘ᆅ࡛῝้ࡋ࡚࠾ࡾࠊ⏘ᆅࡁ࡞⿕ᐖࢆ࠼࡚࠸ࡿ 㸦Weber ࡽ㸪2006㸧ࠋ୍᪉ࠊᚷᆅ⌧㇟ࡢⓎ⏕ᅉᏊࡣᐖࡸࣞࣟࣃࢩ࣮࠸ࡗࡓ⏕≀࣭Ꮫⓗᅉ Ꮚࡀุ᫂ࡋ࡚࠸ࡿࡀᮍࡔ᫂࡞Ⅼࡣከࡃࠊ㜵㝖ᑐ⟇ࡢ㛤Ⓨࡶ㐜ᘏࡋ࡚࠸ࡿࠋ ᚷᆅ⌧㇟࠾ࡅࡿ⏕≀ⓗᅉᏊࡋ࡚ࡣࣇࢨ࣒ࣜ࢘⳦ࡼࡿᐖࡀุ᫂ࡋ࡚࠾ࡾ㸦Vujanobic ࡽ㸪 2006㸧ࠊ᪥ᮏᅜෆ࡛ࡣ Fusarium oxysporum f.sp. asparagi ཬࡧ F. proliferatum ࡀඃ༨✀࡛࠶ࡾࠊࡑࢀ
ࡽࡼࡿ❧ᯤཬࡧᰴ⭉㸦Fusarium crown and root rot㸪FCRR㸧ࡀ࡞ࣇࢨ࣒ࣜ࢘⪃࠼ࡽࢀ
࡚࠸ࡿ㸦Nahiyan ࡽ㸪2008㸧ࠋࡋࡋࠊᐖ㜵㝖࣭᳜≀⏕⫱ᨵၿࢆ㍈ࡋࡓᚷᆅ⌧㇟ᑐ⟇ἲࡢ☜❧
ࡘ࠸࡚ࡣ◊✲ࡀᑡ࡞ࡃࠊᅜෆ࡛ࡢ᳨ウࡣᅜእẚ㐜ᘏࡋ࡚࠸ࡿࠋ
㏆ᖺ࡛ࡣࠊ⏕ែ⣔ಖࢆ⪃៖ࡋࡓᣢ⥆ྍ⬟࡞㎰ᴗࡀᶍ⣴ࡉࢀࡿ୰ࠊᏛ⸆ࡸᏛ⫧ᩱ➼ࡢ௦ ᭰ᡭἲࡋ࡚ࠊᅵተᚤ⏕≀ࢆ⏝࠸࡚ᅵተᐖᢚไࢆᅗࡿ⏕≀ⓗ㜵㝖ἲࡀὀ┠ࡉࢀ࡚࠸ࡿࠋࢳࢦ
ⴎ㯤ࡢࡼ࠺࡞ Fusarium ᐖᑐࡋ࡚ࡣࠊ㠀ཎᛶ Fusarium oxysporum ࡸ AMF㸦arbuscular
mycorrhizal fungi㸸⳦᰿⳦㸧➼ࡼࡿ㜵㝖ࡢヨࡳࡀሗ࿌ࡉࢀ࡚࠸ࡿ㸦Damicone and Manning, 1982; Schneider, 1984; ᑠᕝ࣭㥖⏣, 1984; Paulitz ࡽ㸪1987; Park ࡽ㸪1988; ᡭሯ࣭∾㔝, 1991; Akköprü and Demir, 2006㸧ࠋAMF ࡢሙྜࠊ⪏ᛶࡢࠊ᳜≀య⏕⫱ಁ㐍ຠᯝ㸦Matsubara ࡽ㸪1995, 2004; Wu ࡽ㸪2006㸧ཬࡧ㔞ࡢྥୖ㸦Ozgonen and Erkiric, 2007㸧ࠊ㧗 ࠊ⇱ࠊሷᐖࠊ㔜㔠ᒓ➼ࡢྛ✀⎔ቃ ࢫࢺࣞࢫ⪏ᛶࡀᚓࡽࢀࡿࡓࡵ㸦Hirrel and Gerdemann, 1980; El-Kherbawy ࡽ㸪1989; Ruiz-Lozano ࡽ㸪 1996; Al-Karaki, 2000; Matsubara ࡽ㸪2000㸧ࠊ⥲ྜⓗ࡞᳜≀య⏕⫱ᨵၿἲࡢほⅬࡽᚤ⏕≀㈨ᮦ
ࡋ࡚᭷ᮃ⪃࠼ࡽࢀࡿࠋAMF ࡣ⣒≧⳦ࡢ୍✀࡛ࠊ㝣ୖ᳜≀ࡢ㒊ศឤᰁࡋࠊឤᰁ᰿ࡢ⓶ᒙ⣽
⬊㛫㝽ࡸ⣽⬊ෆᶞᯞ≧యࡸᄞ≧యࡤࢀࡿඹ⏕ჾᐁࢆᙧᡂࡍࡿࠋAMF ࡣᶞᯞ≧యࢆ㏻ࡌ࡚ᐟ
᳜≀ࡽ᭷ᶵ≀ࢆཷࡅྲྀࡾࠊእ⏕⳦⣒ࡼࡗ࡚ࣜࣥࢆᅵተ୰ࡽ྾ࡋࠊᐟ౪⤥ࡍࡿࡇ
ࡼࡗ࡚᳜≀య⏕㛗ಁ㐍ຠᯝࢆࡶࡓࡽࡋ࡚࠸ࡿ㸦Marschner and Dell㸪1994; Smith ࡽ㸪2003㸧ࠋ
17
ࡼࡿFCRR ⪏ᛶㄏᑟࡢྍ⬟ᛶࡀ♧၀ࡉࢀ࡚࠾ࡾ㸦Reid ࡽ㸪2001㸹Elmer㸪2002㸧ࠊᮏ◊✲ᐊ࠾
࠸࡚ࡶࠊNaCl ࡼࡿ❧ᯤ⪏ᛶㄏᑟࡢྍ⬟ᛶࡀ♧၀ࡉࢀ࡚࠸ࡿ(Okada and Matsubara㸪2012)ࠋ୍
᪉ࠊࣇࢨ࣒ࣜ࢘ᑐࡋ࡚㠀ཎᛶF. oxysporum ࡀⓎᢚไຠᯝࢆ᭷ࡍࡿࡇࡘ࠸࡚ࡣከࡃࡢ ሗ࿌ࡀ࠶ࡿ ᑠᕝ࣭㥖⏣1984 ࠋࡇࢀ㛵㐃ࡋࠊNahiyan ࡽ(2010)ࡣࠊࢫࣃࣛ࢞ࢫ࠾ࡅࡿ AMFࠊ 㠀ཎᛶࣇࢨ࣒ࣜ࢘⳦ࡼࡿ⏕⫱ಁ㐍ຠᯝⓎᢚไຠᯝࢆ♧၀ࡋ࡚࠸ࡿ(Nahiyan ࡽ㸪2010)ࠋ ࡋࡋࠊࡇࢀࡽࡢ⏕≀ⓗ࣭Ꮫⓗᡭἲࡼࡿ⪏ᛶㄏᑟ࠾ࡅࡿㄏᑟᢠᛶࡢ㛵ཬࡧㄏᑟᢠ ᛶᶵᵓ㛵ࡋ࡚ࡣ᫂ࡽࡉࢀ࡚࠸࡞࠸ࠋ ୍᪉ࠊ᳜≀య࡛ࡣගྜᡂࢆ⾜࠺㝿㓟⣲ࢆⓎ⏕ࡋࠊ㐣࡞㟁ᏊࡀᏑᅾࡍࡿ㓟⣲㟁Ꮚࡢཷᐜయ ࡋ࡚᭷ẘ࡞ࢫ࣮ࣃ࣮࢜࢟ࢩࢻ㸦O2-㸧ࡸ㐣㓟Ỉ⣲㸦H2O2㸧ࢆࡣࡌࡵࡋࡓࠊࣄࢻࣟ࢟ࢩࣛࢪ࢝
ࣝ㸦㺃OH㸧ࠊ୍㔜㡯㓟⣲㸦1O2㸧࡞ࡢάᛶ㓟⣲ࢆ⏕ࡌࡿ(Foyer ࡽ㸪1994; Asada, 1999)ࠋࡑࡢࡓࡵࠊ
᳜≀య࡛ࡣࢫ࣮ࣃ࣮࢜࢟ࢩࢻࢪࢫ࣒ࢱ࣮ࢮ㸦SOD㸧ࡸࢫࢥࣝࣅࣥ㓟࣌ࣝ࢜࢟ࢩࢲ࣮ࢮ㸦APX㸧ࠊ ࢝ࢱ࣮ࣛࢮ㸦CAT㸧࠸ࡗࡓᢠ㓟㓝⣲ࡸ࣏ࣜࣇ࢙ࣀ࣮ࣝࡸࢫࢥࣝࣅࣥ㓟➼௦⾲ࡉࢀࡿᢠ㓟 ≀㉁ࡼࡿᢠ㓟ᶵ⬟ࢆάᛶࡉࡏࠊ㏻ᖖཬࡧࢫࢺࣞࢫ᮲௳ୗ࡛Ⓨ⏕ࡍࡿάᛶ㓟⣲ࢆ㎿㏿㝖 ཤࡋࠊ௦ㅰࢆ⥔ᣢࡋ࡚࠸ࡿ㸦ὸ⏣1990;┿㔝࣭ὸ⏣1999;Ụࡽ 2002㸧ࠋࡋࡋࠊάᛶ㓟⣲ ࡢ⏕ᡂᢚไࡸᾘཤࡀ༑ศᶵ⬟ࡋ࡞࠸ሙྜࠊ⺮ⓑ㉁ᦆയࠊDNA ᦆയࠊ⭷ᦆയࠊගྜᡂ㜼ᐖࠊ⪁ࠊ ⤌⧊Ṛ࡞ᵝࠎ࡞⏕⌮ⓗ㞀ᐖࡸᝈࡀ⏕ࡌࡿࡇࡀ᫂ࡽ࡞ࡗ࡚࠸ࡿ㸦Kaiser, 1976; Dhindsa
ࡽ㸪1981; McRae and Thompson, 1983; Powles, 1984; Panavas and Rubinstein㸪1998; Asada, 1999; ᮤࡽ
2001; Sohal ࡽ㸪2002㸧ࠋࡲࡓࠊᙉගࡸ㧗 ࠊప ࠊ⇱ࠊሷ➼ࡢࢫࢺࣞࢫ⎔ቃୗ࡛ࡣάᛶ㓟⣲Ⓨ
⏕ࡀຓ㛗ࡉࢀࡿࡀࠊྠSOD ࡸ APX ࠸ࡗࡓᢠ㓟㓝⣲άᛶࡀ㧗ࡲࡗࡓࡢሗ࿌ࡀ࠶ࡿ㸦Foster
and Hess, 1982; Feierabend ࡽ㸪1992; O’kane ࡽ㸪1996; Fadzillah ࡽ㸪1996; Asada, 1999; Kang and
Saltvait, 2002;Gulen and Eris, 2004㸧ࠋ୍᪉ࠊཎ⳦ᑐࡍࡿ᳜≀యࡢ⏕⌮ⓗᛂࡘ࠸࡚ࡣࠊࢪࣕ
࢞ࣔሢⱼษ᩿㠃⤌⧊㠀ぶᛶࡢ⳦⳦యቨ࢚ࣜࢩࢱ࣮㸦HWC㸧ࢆฎ⌮ࡋࡓሙྜ O2-ࡢ⏕
ᡂࡀㄆࡵࡽࢀ㸦Doke, 1983㸧ࠊࢿ⛉ࡸࢼࢫ⛉ࠊ࣐࣓⛉࠸ࡗࡓᵝࠎ࡞᳜≀✀࡛ࡶཎ⳦ࡢឤᰁࡸ
࢚ࣜࢩࢱ࣮ฎ⌮ࡼࡿ O2-ࡸH2O2ࡢ⏕ᡂࡀ᫂ࡽ࡞ࡗ࡚࠸ࡿ㸦Apostol ࡽ㸪1989;ᕝ࣭㐨ᐙ,
18
ရ✀࠾࠸࡚ SOD άᛶࡀపୗࡋࠊO2-✚㔞ࡀከࡃ࡞ࡗࡓ⤖ᯝࠊ㐣ᩄឤᛂࢆㄏᑟࡋࡓ࠸࠺ሗ
࿌ࡀ࠶ࡿ㸦Zacheo ࡽ㸪1988㸧ࠋࡇࢀࡽࡣᑐ↷ⓗࠊࢺ࣐ࢺࡢཎ⳦ Botrytis cinerea ᥋✀ᚋ
SOD άᛶࡢቑࡀࡳࡽࢀ㸦Kuźniak and Skłodowska, 2004㸧ࠊࣥࢤࣥࡉࡧࡸࢳࢦ⺬ࡢ┠➼
࠾࠸࡚ࡣࠊᢠᛶရ✀࡛ࡣ⨯ᛶရ✀ࡼࡾࡶཎ⳦᥋✀ᚋࡢ᪩ᮇSOD άᛶࡀቑࡍࡿࡇࡀሗ
࿌ࡉࢀ࡚࠸ࡿ㸦Buonaurio ࡽ㸪1987; Ehsani-Moghaddam ࡽ㸪2006; Sahoo ࡽ㸪2007㸧ࠋ୍᪉ࠊⓎ
ᢠ㓟≀㉁ࡢ㛵㐃ࡘ࠸࡚ࡣࠊࢫࢥࣝࣅࣥ㓟ྵ㔞ࡀ㔝ᛶᰴẚ㍑ࡋ࡚30%ప࠸ࢩࣟࢾࢼࢬ ࢼኚ␗యཎ⳦ࢆ᥋✀ࡍࡿࠊ㜵ᚚࢱࣥࣃࢡ㉁࡛࠶ࡿβࢢࣝ࢝ࢼ࣮ࢮཬࡧ࢟ࢳࢼ࣮ࢮࡸ PR-1ࠊ PR-2 ࠸ࡗࡓ PR㸦pathogenesis-related㸧ࢱࣥࣃࢡ㉁࡞ࡢ㑇ఏᏊࡢⓎ⌧ࡀቑຍࡋࠊᐖࡀ㍍ῶࡉ ࢀࡓࡢሗ࿌ࡀ࠶ࡿ㸦Pastori ࡽ㸪2003㸧ࠋࡉࡽࠊࡇࡢኚ␗యࢫࢥࣝࣅࣥ㓟ࢆᢞࡍࡿࠊ ㏫PR-1 ࡢⓎ⌧ࡀῶᑡࡋࡓࡇࡽ㸦Pastori ࡽ㸪2003㸧ࠊࢫࢥࣝࣅࣥ㓟ࡢῶᑡࡀ㜵ᚚ㑇ఏᏊⓎ ⌧㛵㐃ࡋ࡚࠸ࡿྍ⬟ᛶࡶ⪃࠼ࡽࢀ࡚࠸ࡿࠋࡲࡓࠊࢫࢥࣝࣅࣥ㓟ࡣᢠ㓟≀㉁࡛࠶ࡿྠࠊ ࢫࢥࣝࣅࣥ㓟࣌ࣝ࢜࢟ࢩࢲ࣮ࢮ㸦APX㸧ࡢᇶ㉁࡛ࡶ࠶ࡿࡓࡵࠊAPX άᛶᙳ㡪ࢆ࠼ࡿࡇࡀ ▱ࡽࢀ࡚࠸ࡿ㸦㔜ᒸ1999㸧ࠋࡇࡢࡼ࠺ࠊ⪏ᛶ᳜≀యࡢᢠ㓟ᶵ⬟ࡣ┤᥋ⓗཬࡧ㛫᥋ⓗ࡞㛵 ಀࡀ࠶ࡿࡇࡀ♧၀ࡉࢀ࡚࠸ࡿࠋ ᳜≀యࡢᢠ㓟ᶵ⬟ኚືAMF ඹ⏕ࡢ㛵㐃ࡘ࠸࡚ࡣࠊ࢚ࣥࢻ࡛࢘ AMF ᥋✀ 23 ᪥ᚋཬࡧ
30 ᪥ᚋ SOD άᛶࡀ㧗ࡲࡾ(Arines ࡽ㸪1994)ࠊࢱࣂࢥ࠾࠸࡚ࡣ᥋✀ 5 ᪥ᚋ APX CAT ࡀቑ
ࡋࡓࡀ࠶ࡿ㸦Blilou ࡽ㸪2000㸧ࠋࡲࡓࠊ㧗 ࢫࢺࣞࢫ᮲௳⛣⾜ࡉࡏࡓ AMF ඹ⏕ࢳࢦ᳜ ≀య࡛ࡣ㧗 ⪏ᛶࡀࡳࡽࢀࠊ㧗 ⛣⾜5 ᪥ᚋཬࡧ 10 ᪥ᚋ࡛ࡣࠊSOD ཬࡧ APX άᛶࡢቑ࣭⥔ᣢ ࡀㄆࡵࡽࢀ࡚࠸ࡿ㸦Li ࡽ㸪2008㸧ࠋࡉࡽࠊࢫࣃࣛ࢞ࢫཬࡧࢳࢦ࠾࠸࡚ AMF ඹ⏕య࡛ࡣ 㧗 ⛣⾜ᚋᢠ㓟㓝⣲ཬࡧᢠ㓟≀㉁ྵ㔞ࡀቑ࣭⥔ᣢࡉࢀࡿ⤖ᯝࡀᚓࡽࢀ࡚࠸ࡿ㸦ᐑ⬥ 2007㸧ࠋࡇࡢࡼ࠺ࠊᩘ✀᳜≀࠾࠸࡚ $0) ඹ⏕ࡼࡾᢠ㓟㓝⣲άᛶࡀ㧗ࡲࡿࡇࡀ♧၀ࡉࢀ࡚ ࠸ࡿࡀࠊAMFࠊ㠀ཎᛶࣇࢨ࣒ࣜ࢘⳦ཬࡧ NaCl ฎ⌮ࡼࡿࢫࣃࣛ࢞ࢫ࠾ࡅࡿཎ⳦᥋✀๓ ᚋࡢᢠ㓟ᶵ⬟ኚືཬࡧㄏᑟᢠᛶࡢ㛵㐃ࡘ࠸࡚ࡣ᫂Ⅼࡀከ࠸ࠋ ᮏ◊✲࡛ࡣࠊAMF ࢆ⏝ࡋࡓࢫࣃࣛ࢞ࢫࡢ⥲ྜⓗ᳜≀ᨵၿἲࢆ☜❧ࡍࡿࡓࡵࠊᐇ⏝ရ✀࡛ ࠶ࡿࢢ࣮ࣜࣥ⣔ 3 ရ✀⣸⣔ 2 ရ✀࠾ࡅࡿ AMF ᥋✀ࡼࡿ⪏ᛶㄏᑟࡢရ✀㛫ᕪ␗ࡘ࠸࡚
19
᳨ウࡋࡓࠋࡲࡓࠊsplit root system ἲࢆ⏝࠸࡚ AMFࠊ㠀ཎᛶࣇࢨ࣒ࣜ࢘⳦ཬࡧ NaCl ࡼࡿ❧ᯤ
ㄏᑟᢠᛶࡢ᳨ドࢆ⾜࠺ࡶࠊࢫࣃࣛ࢞ࢫ࡛ࡢ⪏ᛶᶵᵓゎ᫂ࡢ୍⎔ࡋ࡚ࠊ❧ᯤ⳦ ᥋✀๓ᚋ࠾ࡅࡿᢠ㓟㓝⣲ࡸᢠ㓟≀㉁ࠊDPPH ࣛࢪ࢝ࣝᾘཤ⬟➼ࢆㄪᰝࡋࠊAMFࠊ㠀ཎᛶ ࣇࢨ࣒ࣜ࢘⳦ཬࡧNaCl ࡼࡿࢫࣃࣛ࢞ࢫ࠾ࡅࡿᢠ㓟ᶵ⬟ཬࡧ SOD ࢯࢨ࣒ኚື⪏ ᛶࡢ㛵㐃ᛶࢆㄪᰝࡋࡓࠋ
20 ᮦᩱཬࡧ᪉ἲ ➨1 ⠇ ࢫࣃࣛ࢞ࢫ࡛ࡢ AMF ࡼࡿ❧ᯤ⪏ᛶㄏᑟࡢရ✀㛫ᕪ 1. AMF ᥋✀ ࣮࢜ࢺࢡ࣮ࣞࣈ(120Υ㸪1.2kg/cm2㸪20 ศ)ࡋࡓᕷ㈍⫱ⱑᇵᅵ㸦SM-2㸪ᥬᩫᕝᕤᴗᰴᘧ♫㸧 ࢆࣂࢵࢺ㸦13.5×27.0×15.5cm㸧ワࡵࠊࢫࣃࣛ࢞ࢫࡢࢢ࣮ࣜࣥ⣔ 3 ရ✀㸦‘࢙࣒࢘ࣝ࢝’㸪͂ࢫ࣮ ࣃ࣮࢙࣒࢘ࣝ࢝̓㸪͂㞝࢞ࣜࣂ࣮̓ 㸧⣸⣔ 2 ရ✀㸦͂ࣂ࣮࢞ࣥࢹ࣮̓㸪͂‶⣸̓㸧ࢆ
✀ࡋࡓࠋࡑࡢ㝿ࠊAMF 㹙Glomus intraradices㸦Gi㸧㹛ࢆ᥋✀㸦2g /ಶయ㸧ࡋࠊᑐ↷༊ࡣ࣮࢜ࢺࢡ
࣮ࣞࣈࡋࡓAMF ᥋✀≀ࢆฎ⌮ࡋࡓࠋ౪ヨಶయᩘࡣྛヨ㦂༊ 20 ಶయࡋࠊ⫧ࡣ⦆ຠᛶ⫧ᩱ㹙ࣟ ࣥࢢࢺ࣮ࢱࣝ70 ᪥ࢱࣉ㸦N : P : K㸻13 : 11 : 13㸧㸪᪫ᡂᰴᘧ♫㹛ࢆ⏝㸦0.1g/ಶయ㸧ࡋࠊᒱ 㜧Ꮫࡢ࢞ࣛࢫ ᐊෆ࡛㐺ᐅ₤Ỉࢆ⾜࠸ࠊ⮬↛᪥㛗ୗ࡛⫱ⱑࡋࡓࠋ 2.⏕⫱ㄪᰝ AMF ᥋✀ 10 㐌㛫ᚋ᳜≀యࢆ᥇ྲྀࡋ࡚ᆅୖ㒊࠾ࡼࡧᆅୗ㒊≀㔜ࢆㄪᰝࡋࡓࠋㄪᰝಶయࡣ 10 ಶయࡋࡓࠋ 3.ཎ⳦᥋✀
PDA ᇵᆅࢆ⏝࠸࡚⣧⢋ᇵ㣴ࡋࡓࢫࣃࣛ࢞ࢫ❧ᯤ⳦㹙Fusarium oxysporum f. sp. asparagi㸦Foa : MAFF 305556㸧㹛ࡢ⳦ࡑ࠺ࢆ potato sucrose㸦PS㸧ᇵᆅΰྜࡋࠊ25Υࠊᬯᡤ࡛ᇵ㣴ࡋࡓࠋࡑࡢᚋࠊ
Foa ศ⏕⬊Ꮚᠱ⃮ᾮࢆ 105⬊ᏊȀㄪᩚࡋࠊAMF ᥋✀ 10 㐌㛫ᚋࡢࢫࣃࣛ࢞ࢫᐇ⏕᰿㒊᥋✀ 㸦50ml/ಶయ㸧ࡋࡓࠋཎ⳦᥋✀ᚋࡣ࢞ࣛࢫ ᐊෆ࡛㐺ᐅ₤Ỉࢆ⾜࠸㝸㞳⫱ⱑࡋࡓࠋ 5.Ⓨㄪᰝ ཎ⳦᥋✀6 㐌㛫ᚋࠊ᳜≀యࢆ᥇ྲྀࡋ࡚Ⓨㄪᰝࢆ⾜ࡗࡓࠋㄪᰝಶయᩘࡣྛヨ㦂༊ 10 ಶయࡋࠊ Ⓨ⋡࠾ࡼࡧⓎ⛬ᗘࢆྛಶయࡢ㈓ⶶ᰿ᩘᑐࡍࡿ⨯᰿ᩘࡢྜ࡛ồࡵࠊ6 ẁ㝵タᐃࡋࡓ Ⓨᣦᩘ࡛ホ౯ࡋࡓࠋࡲࡓࠊⓎᗘࢆ௨ୗࡢᘧࡼࡾ⟬ฟࡋࡓࠋ
21 Ⓨᣦᩘ
ྛಶయࡢ㈓ⶶ᰿ᩘᑐࡍࡿ⨯᰿ᩘࡢྜ
0㸸0% 1㸸~20㸣 2㸸20~40㸣 3㸸40~60% 4㸸60~80% 5㸸80~100%
➨2 ⠇ Split root system ἲࡼࡿࢫࣃࣛ࢞ࢫ࡛ࡢ⏕≀࣭Ꮫⓗᡭἲࡼࡿㄏᑟᢠᛶホ౯ཬࡧ
ᢠ㓟 ᶵ⬟ኚືゎᯒ
2-1 Split root systemࡼࡿࢫࣃࣛ࢞ࢫ❧ᯤࡢㄏᑟᢠᛶホ౯ 1㸬ࢫࣃࣛ࢞ࢫ✀
࢜ ࣮ ࢺ ࢡ ࣞ ࣮ ࣈ ࡋ ࡓ ᕷ ㈍ ⫱ ⱑ ᅵ 㸦 ࢭ ࣝ ᇵ ᅵ TM 㸫 1 㸹 ࢱ ࢟ ✀ ⱑ ᰴ ᘧ ♫ 㸧 ࢆ ࣂ ࢵ ࢺ
㸦19.0×33.5×15.5cm㸧ワࡵࠊࢫࣃࣛ࢞ࢫ㸦Asparagus officinalis L., ‘Welcome’㸧ࢆ✀ࡋࠊ⫧
ࡣ⦆ຠᛶ⫧ᩱ㸦ࣟࣥࢢࢺ࣮ࢱࣝ 180 ᪥ࢱࣉ㸪N:P:K㸻13:11:13㸪ࢳࢵࢯ᪫ᰴᘧ♫㸧ࢆ⏝ࡋࠊ
ᐊෆ࡛㐺ᐅ₤Ỉ㸦⣙50mlᅇ㸪2 ᅇday㸧ࢆ⾜࠸⫱ⱑࡋࡓࠋ
2㸬AMF, 㠀ཎᛶࣇࢨ࣒ࣜ࢘⳦ཬࡧ NaCl ฎ⌮
➨6 ᅗ♧ࡋࡓࡼ࠺✀ 8 㐌㛫ᚋ᰿⣔ࢆ 2 ศࡋࠊฎ⌮᰿㸦R㸧ཬࡧ↓ฎ⌮᰿㸦L㸧ࢆタᐃ
ࡋࡓ ௨ୗ split root system ⛠ࡍ➨ 7 ᅗ㸧ࠋྛฎ⌮༊ࡢᐇ⏕ࡣ 20 ಶయࡋࠊAMF㹙Glomus
intraradices㸦Gi㸧㸹ฟගࢢࣜᰴᘧ♫ࡼࡾศㆡ㹛ࢆ✀ 8 㐌㛫ᚋ R ᥋✀ࡋࡓ㸦2gಶయ㸧ࠋ
AMF ᥋✀ 8 㐌㛫ᚋࡽ 1 㐌㛫ẖ NaCl Ỉ⁐ᾮ㸦50mM㸧ࢆ R 1 ಶయᙜࡓࡾ 10ml ฎ⌮ࡋࡓࠋ୍ ᪉ࠊPotato dextrose agar㸦PDA㸧ᇵᆅࢆ⏝࠸࡚⣧⢋ᇵ㣴㸦25Υ㸪14 ᪥㛫㸪ᬯᡤ㸧ࡋࡓ㠀ཎᛶࣇࢨ
࣒ࣜ࢘⳦㹙Non-pathogenic Fusarium oxysporum㸦NPFO, NF1㸧㹛ࡢ⳦ࡑ࠺ࢆ potato sucrose㸦PS㸧ᇵ
ᆅΰྜࡋࠊ25Υࠊᬯᡤ࡛ 2 㐌㛫ᇵ㣴ࡋࡓࠋࡑࡢᚋࠊNPFO ࡢศ⏕⬊Ꮚᠱ⃮ᾮࢆ 105⬊Ꮚ/ml ㄪ
ᩚࡋࠊAMF ᥋✀ 19 㐌㛫ᚋ R ᅵተ₤ὀ᥋✀㸦50mlಶయ㸧ࡋࡓࠋ
Ⓨᗘ 㸻 㸻 ∑㸦ಶయᩘ× Ⓨᣦᩘ㸧 × 100×
22 3㸬❧ᯤ⳦᥋✀
2 ྠᵝࡢ᪉ἲ࡛ᇵ㣴ࡋࡓࢫࣃࣛ࢞ࢫ❧ᯤ⳦㹙F. oxysporum f. sp. asparagi, MAFF305556
㸦Foa㸧㹛ࡢศ⏕⬊Ꮚᠱ⃮ᾮࢆ 106⬊Ꮚ/ml ㄪᩚࡋࠊAMF ᥋✀ 21 㐌㛫ᚋ࡚ࡢ᰿ᇦᅵተ₤ὀ ᥋✀㸦50mlಶయ㸧ࡋࡓࠋཎ⳦᥋✀ᚋࡣ ᐊෆ࡛㐺ᐅ₤Ỉࢆ⾜࠸㝸㞳⫱ⱑࡋࡓࠋ 4㸬⏕⫱ㄪᰝ AMF ᥋✀ 21 㐌㛫ᚋࡢᆅୖ㒊ᆅୗ㒊ཬࡧཎ⳦᥋✀ 4 㐌㛫ᚋࡢᆅୗ㒊ࡢ᳜≀యࢆ᥇ྲྀࡋࠊ ≀㔜ࢆㄪᰝࡋࡓࠋㄪᰝಶయᩘࡣྛ༊5 ಶయࡋࡓࠋ 5㸬᰿⤌⧊ෆ࠾ࡅࡿ AMF ឤᰁ≧ែࡢㄪᰝ
Phillips and Hayman㸦1970㸧ࡢ᪉ἲᚑ࠸ࠊAMF ᥋✀ 21 㐌㛫ᚋཬࡧཎ⳦᥋✀ 4 㐌㛫ᚋࡢྛᐇ
⏕ࡢ྾᰿ࢆ10㸣KOH ⁐ᾮᾐₕࡋࡓ≧ែ࡛࣮࢜ࢺࢡ࣮ࣞࣈ㸦120Υ㸪1.2kg⁄cm2㸪15 ศ㸧ࡋࡓࠋ ࡑࡢᚋࠊࢺࣜࣃࣥࣈ࣮ࣝ⁐ᾮ㸦ࢢࣜࢭࣜࣥ50ml㸪ங㓟 50ml㸪␃Ỉ 50ml㸪ࢺࣜࣃࣥࣈ࣮ࣝ1g ࢆΰྜࡋࠊ70%࢚ࢱࣀ࣮࡛ࣝ 500ml ࣓ࢫࢵࣉ㸧ࡼࡾᰁⰍࡋࡓࠋᰁⰍࡉࢀࡓ᰿ࢆ 0.5㹼1.0cm ࡢ㛗ࡉ⣽᩿ࡋࠊ᰿ษ∦ࢆࢫࣛࢻࢢࣛࢫୖࡢࡏࠊ࢝ࣂ࣮ࢢࣛࢫ࡛そ࠸ࠊගᏛ㢧ᚤ㙾ࢆ⏝࠸࡚ ឤᰁ⋡ࡘ࠸࡚ㄪᰝࡋࡓࠋឤᰁ⋡ࡣࠊ྾᰿ษ∦ᩘᑐࡍࡿ AMF ࡀឤᰁࡋࡓ྾᰿ษ∦ᩘࡢ ྜࡋࠊྛ᥋✀༊ࡶࡣ3 ࡋࡓࠋ 6㸬Ⓨホ౯ ཎ⳦᥋✀4 㐌㛫ᚋࠊ᳜≀యࢆ᥇ྲྀࡋ࡚ྛ༊ 10 ಶయࡘ࠸࡚Ⓨㄪᰝࢆ⾜ࡗࡓࠋⓎ⛬ᗘࡋ ࡚ࠊྛಶయࡢ㈓ⶶ᰿ᩘᑐࡍࡿ⨯᰿ᩘࡢྜࢆồࡵࠊ6 ẁ㝵࡛ࣞ࣋ࣝホ౯ࡋࠊⓎᣦᩘࢆ௨ ୗࡢᘧࡼࡾ⟬ฟࡋࡓࠋ Ⓨࣞ࣋ࣝ 0㸸0% 1㸸20%ᮍ‶ 2㸸20%௨ୖ 40%ᮍ‶ 3㸸40%௨ୖ 60%ᮍ‶ 4㸸60%௨ୖ 80%ᮍ‶ 5㸸80%௨ୖ
23 Ⓨᣦᩘ㸻 × 100 2-2 ᢠ㓟ᶵ⬟ኚືゎᯒ 1㸬ヨᩱస〇 2-1 ࡛ᚓࡽࢀࡓࠊAMF ᥋✀ 21 㐌㛫ᚋཬࡧཎ⳦᥋✀ 4 㐌㛫ᚋࡢࢫࣃࣛ࢞ࢫ᳜≀యࢆࡑࢀࡒࢀ ⱼࠊ㈓ⶶ᰿㸦ฎ⌮᰿ࠊ↓ฎ⌮᰿㸧㢮ูࡋࠊᾮయ❅⣲࡛⤖ࡋࡓࠋ 2㸬SOD άᛶ
SOD άᛶࡢ ᐃࡣ Beauchamp and Fridovich㸦1971㸧ࡢ᪉ἲ‽ࡌ࡚⾜ࡗࡓ㸦➨ 8 ᅗ㸧ࠋ⤖ヨ
ᩱ0.1g 50mM ࣜࣥ㓟⦆⾪ᾮ㸦pH7.0㸧3.0ml ࢆῧຍࡋࠊ෭༷ࡋࡓங㖊ࢆ⏝࠸࡚☻○ᚋࠊ㐲ᚰศ㞳 ᶵ㸦EF㸫1300㸪ࢺ࣑࣮ᕤᴗᰴᘧ♫㸧࡛㧗㏿㐲ᚰ㸦13,000rpm㸪5Υ㸪5 ศ㸧ࡋࡓୖΎࢆ⢒㓝⣲ᾮ ࡋࡓࠋ50mM Ⅳ㓟ࢼࢺ࣒ࣜ࢘⦆⾪ᾮ㸦pH10.2㸧2.3mlࠊ10mM NBT 0.1mlࠊ4.0mM ࢟ࢧࣥࢳࣥ 0.1mlࠊ 3.0mM EDTA 0.1mlࠊ0.15%㸦w/v㸧BSA㸦∵⾑Ύࣝࣈ࣑ࣥ㸧0.1ml ࡢ㡰ΰྜࡋࡓᚋࠊ⢒㓝⣲ᾮ 0.1ml ࢆຍ࠼ࡓࠋࡑࡢΰྜᾮ 6.0 units/ml ࢟ࢧࣥࢳࣥ࢜࢟ࢩࢱ࣮ࢮ㸦100 ಸᕼ㔘ᾮ㸧0.1ml ࢆῧຍ ࡋ࡚ᛂࢆ㛤ጞࡉࡏࡓࠋ30Υࠊᬯᡤ࡛ 30 ศ㛫࣮ࣥ࢟ࣗ࣋ࢺᚋࠊ14mM CuCl2 0.2ml ࡢῧຍࡼ ࡗ࡚ᛂࢆṆࡉࡏࠊ⏕ᡂࡋࡓ㟷Ⰽࣇ࢛࣐ࣝࢨࣥࢆ560nm ࡢἼ㛗࡛ศගගᗘィ㸦U㸫1900㸪᪥❧ ࣁࢸࢡࣀࣟࢪ࣮ࢬᰴᘧ♫㸧ࡼࡾ ᐃࡋࡓࠋࡲࡓࠊᑐ↷྾ගᗘࢆ 50%ᢚไࡍࡿ SOD άᛶࢆ 1unit ࡋࡓࠋ 3㸬DPPH ࣛࢪ࢝ࣝᤕᤊ⬟ DPPH ࣛࢪ࢝ࣝᤕᤊ⬟ࡢ ᐃࡣᑠᯘ㸦1999㸧ࡢ᪉ἲ‽ࡌ࡚⾜ࡗࡓ㸦➨ 9 ᅗ㸧ࠋ⢭⛗ࡋࡓ⤖ヨ ᩱ0.1g 80㸣࢚ࢱࣀ࣮ࣝ 3ml ࢆຍ࠼࡚☻○ࡋࠊ㧗㏿㐲ᚰ㸦13,000rpm㸪5Υ㸪5 ศ㸧ᚋࠊୖΎࢆศ ᯒヨᩱࡋࡓࠋ⥆࠸࡚ヨ㦂⟶400μM DPPH ⁐ᾮࠊ0.2M MES ⦆⾪ᾮ㸦pH6.0㸧ࠊ20%࢚ࢱࣀ࣮ࣝ ࢆྛ0.9ml ࡎࡘຍ࠼࡚ΰྜࡋࠊ80%࢚ࢱࣀ࣮ࣝ 0.75mlࠊศᯒヨᩱ 0.15ml ࢆຍ࠼࡚ 30Υࠊᬯᡤ࡛ 30 Ȋ㸦ಶయᩘⓎࣞ࣋ࣝ㸧 ౪ヨಶయᩘ5㸦᭱㔜ᗘࣞ࣋ࣝ㸧
24 ศ㛫㟼⨨ࡋࡓࠋࡑࡢ㝿ࠊศᯒヨᩱࡢ௦ࢃࡾ80%࢚ࢱࣀ࣮ࣝ 0.15ml ຍ࠼ࡓࡶࡢࢆࣈࣛࣥࢡࡋࠊ ศගගᗘィࡼࡾ520nm ࠾ࡅࡿ྾ගᗘࢆ ᐃࡋࡓࠋ᳨㔞⥺సᡂࡣ Trolox㸦0㹼150μg⁄ml㸧ࢆ⏝ ࠸ࠊヨᩱᾮࡢ ᐃ್ࡽࣈࣛࣥࢡ್ࢆᕪࡋᘬࡁࠊ᳨㔞⥺࡛⟬ฟࡋࡓ್ࢆDPPH ࣛࢪ࢝ࣝᤕᤊ⬟ ࡋࡓࠋ 㸬⥲ࢫࢥࣝࣅࣥ㓟ศᯒ ⥲ࢫࢥࣝࣅࣥ㓟ศᯒࡣࣄࢻࣛࢪࣥẚⰍἲ㸦Roe ࡽ㸪1948㸧‽ࡌ࡚⾜ࡗࡓ㸦➨ 10 ᅗ㸧ࠋ⤖ ヨᩱ0.1g ࢆ⢭⛗ࡋࠊங㖊ࢆ⏝࠸࡚ 5㸣࣓ࢱࣜࣥ㓟㸦࣓ࢱࣜࣥ㓟 5g ␃Ỉ 100ml ࢆຍ࠼࡚⁐ゎ㸧 3ml ࢆῧຍࡋ࡚☻○ࡋࠊ㧗㏿㐲ᚰ㸦13,000rpm㸪5Υ㸪5 ศ㸧ᚋࠊୖΎࢆヨᩱᾮࡋࡓࠋ⥆࠸࡚ヨ㦂 ⟶2 ᮏヨᩱᾮ 0.5ml ࢆධࢀࠊ0.03%DCIP ⁐ᾮ㸦2,6㸫ࢪࢡࣟࣝࣥࢻࣇ࢙ࣀ࣮ࣝࢼࢺ࣒ࣜ࢘ 15mg Ỉ50ml ࢆຍ࠼࡚⁐ゎ㸧1ml ࢆຍ࠼࡚ΰྜࡋࠊヨᩱᾮࢆ⣚Ⰽ࿊Ⰽᚋࠊ2%ࢳ࢜ᒀ⣲㸫5㸣࣓ࢱ ࣜࣥ㓟⁐ᾮ㸦5%࣓ࢱࣜࣥ㓟⁐ᾮ 100ml ࢳ࢜ᒀ⣲ 2g ࢆຍ࠼࡚⁐ゎ㸧0.5ml ࢆຍ࠼ࡓࠋヨᩱᾮࡢ⣚ Ⰽࡀᾘ࠼ࡓᚋࠊ∦᪉ࡢヨ㦂⟶2㸣DNP ⁐ᾮ㸦9N ◲㓟 100ml 2,4㸫ࢪࢽࢺࣟࣇ࢙ࢽࣝࣄࢻࣛࢪ ࣥ2g ࢆῧຍࡋ⁐ゎ㸧0.25ml ࢆῧຍࡋࠊ࢛࣮࢘ࢱ࣮ࣂࢫෆ࡛ 50Υ⥔ᣢࡋࡓࡲࡲ 70 ศᛂࡉࡏࡓࠋ ᛂ⤊ᚋࠊịỈ୰࡛෭༷ࡋ࡞ࡀࡽ 85%◲㓟ࢆᚎࠎῧຍࡋࠊᐊ ࡛ 30 ศ㟼⨨ࡋࡓࠋࡑࡢ㝿ࠊ ᛂ๓2%DNP ⁐ᾮࢆຍ࠼࡞ࡗࡓヨ㦂⟶ 2%DNP ⁐ᾮ 0.25ml ࢆῧຍࡋࠊࡇࢀࢆࣈࣛࣥࢡ ࡋࡓࠋᛂᚋࠊศගගᗘィࢆ⏝࠸࡚Ἴ㛗 520nm ࠾ࡅࡿ྾ගᗘࢆ ᐃࡋࡓࠋ᳨㔞⥺సᡂࡣ L-ࢫࢥࣝࣅࣥ㓟㸦10㹼25μg⁄ml㸧ࢆ⏝࠸ࠊヨᩱᾮࡢ ᐃ್ࡽࣈࣛࣥࢡ್ࢆᕪࡋᘬࡁࠊ᳨㔞⥺ࡽ ⥲ࢫࢥࣝࣅࣥ㓟ྵ㔞ࢆ⟬ฟࡋࡓࠋ 5㸬⥲࣏ࣜࣇ࢙ࣀ࣮ࣝศᯒ
⥲࣏ࣜࣇ࢙ࣀ࣮ࣝศᯒࡣࣇ࢛࣮ࣜࣥࢹࢽࢫἲ㸦Folin and Denis㸪1915㸧‽ࡌ࡚⾜ࡗࡓ㸦➨ 11
ᅗ㸧ࠋ⤖ヨᩱ0.1g ࢆ⢭⛗ࡋࠊị෭ࡋࡓங㖊ࢆ⏝࠸࡚ 80%࢚ࢱࣀ࣮ࣝ 4ml ࢆῧຍࡋ࡚☻○ࡋࠊ㧗
㏿㐲ᚰ㸦13,000rpm㸪5Υ㸪5 ศ㸧ᚋࠊୖΎࢆศᯒ⏝ヨᩱᾮࡋࡓࠋ⥆࠸࡚␃Ỉ 3ml ࢆຍ࠼ࡓヨ
25 ≀10gࠊࣜࣥࣔࣜࣈࢹࣥ㓟㸦12 ࣔࣜࣈࢻ ϭ ࣜࣥ㓟㺃n Ỉ≀㸧2gࠊࣜࣥ㓟 5ml ࢆຍ࠼࡚⁐ゎࡋ ࡓᚋࠊ2 㛫࢛࣮࢘ࢱ࣮ࣂࢫෆ࡛㑏ὶࡋࠊ෭༷ᚋࠊ␃Ỉࢆຍ࠼࡚ 100ml ᐃᐜࡍࡿࠋ㸧0.2ml ࢆ ຍ࠼࡚ᨩᢾᚋࠊ㣬Ⅳ㓟ࢼࢺ࣒ࣜ࢘⁐ᾮ0.4ml ࢆຍ࠼ࠊࡇࢀࢆᐊ ࡛ 30 ศ㟼⨨ࡋࡓࠋࡑࡢ㝿ࠊࣇ ࢛࣮ࣜࣥࢹࢽࢫヨ⸆ࡢ᭰ࢃࡾ␃Ỉ 0.2ml ࢆຍ࠼ࡓࡶࡢࢆㄪ〇ࡋࠊࡇࢀࢆࣈࣛࣥࢡࡋࡓࠋ30 ศᚋࠊศගගᗘィࢆ⏝࠸࡚Ἴ㛗700nm ࠾ࡅࡿ྾ගᗘࢆ ᐃࡋࡓࠋ᳨㔞⥺సᡂࡣࢡ࢙ࣝࢭࢳࣥ 㸦25㹼100μg⁄ml㸧ࢆ⏝࠸ࠊヨᩱᾮࡢ ᐃ್ࡽࣈࣛࣥࢡ್ࢆᕪࡋᘬࡁࠊ᳨㔞⥺ࡽ⥲࣏ࣜࣇ࢙ࣀ ࣮ࣝྵ㔞ࢆ⟬ฟࡋࡓࠋ ➨3 ⠇ AMF ඹ⏕ࢫࣃࣛ࢞ࢫ࠾ࡅࡿ❧ᯤ⪏ᛶㄏᑟඹ⏕≉␗ⓗ SOD ࢯࢨ࣒ኚື 1㸬AMF ᥋✀ ࣮࢜ࢺࢡ࣮ࣞࣈࡋࡓᕷ㈍⫱ⱑᅵ㸦ࢭࣝᇵᅵ TM㸫1㸧ࢆࣂࢵࢺ㸦19.0×33.5×15.5cm㸧ワࡵࠊ
ࢫࣃࣛ࢞ࢫ㸦‘Welcome’㸧ࢆ✀ࡋࡓࠋࡲࡓࠊ✀ AMF2 ⳦✀㹙Glomus intraradices㸦Gi㸧㸪
Gigaspora margarita 㸦GM㸧㸹ࢭࣥࢺࣛࣝ◪Ꮚᰴᘧ♫ࡼࡾศㆡ㹛ࢆ᥋✀㸦2gಶయ㸧ࡋࡓࠋ࡞࠾ࠊ ᑐ↷༊ࡣ࣮࢜ࢺࢡ࣮ࣞࣈࡋࡓAMF ᥋✀≀ࢆฎ⌮ࡋࡓࠋྛฎ⌮༊ࡢᐇ⏕ࡣ 20 ಶయࡋࠊ⫧ࡣ ⦆ຠᛶ⫧ᩱ㸦ࣟࣥࢢࢺ࣮ࢱࣝ180 ᪥ࢱࣉ㸧ࢆ⏝ࡋࠊ ᐊෆ࡛㐺ᐅ₤Ỉࢆ⾜࠸⫱ⱑࡋࡓࠋ 2㸬❧ᯤ⳦᥋✀ ➨2 ⠇ࡢ 2-1 ࡢ 2 ྠᵝࡢ᪉ἲ࡛ᇵ㣴ࡋࡓࢫࣃࣛ࢞ࢫ❧ᯤ⳦㸦Foa; MAFF305556㸧ࡢศ⏕ ⬊Ꮚᠱ⃮ᾮࢆ106⬊Ꮚ/ml ㄪᩚࡋࠊAMF ᥋✀ 8 㐌㛫ᚋࡢࢫࣃࣛ࢞ࢫᐇ⏕᰿㒊ᅵተ℺ὀ᥋✀ ࡋࡓ㸦50ml/ಶయ㸧ࠋཎ⳦᥋✀ᚋࡣ ᐊෆ࡛㐺ᐅ₤Ỉࢆ⾜࠸㝸㞳⫱ⱑࡋࡓࠋ 3㸬⏕⫱ㄪᰝ AMF ᥋✀ 8 㐌㛫ᚋཬࡧཎ⳦᥋✀ 8 㐌㛫ᚋࡢᆅୖ㒊ᆅୗ㒊ࡢ᳜≀యࢆ᥇ྲྀࡋࠊ≀㔜ࢆㄪᰝ ࡋࡓࠋㄪᰝಶయᩘࡣྛ༊5 ಶయࡋࡓࠋ
26 4㸬Ⓨホ౯ ཎ⳦᥋✀8 㐌㛫ᚋࠊ᳜≀యࢆ᥇ྲྀࡋ࡚ྛ༊ 10 ಶయࡘ࠸࡚Ⓨㄪᰝࢆ⾜ࡗࡓࠋⓎホ౯ࡢ᪉ ἲࡣ➨ ⠇ࡢ2-1 ࡢ 6 ྠᵝࡢ᪉ἲ࡛⾜ࡗࡓࠋ 5㸬⳦㔞ࡢ ᐃ ཎ⳦᥋✀8 㐌㛫ᚋࡢ᰿ᅪᅵተཬࡧ㈓ⶶ᰿ࢆ᥇ྲྀࡋࠊྛࢧࣥࣉࣝ 1g ࢆ␃Ỉࢆ⏝࠸࡚ᕼ㔘ࡋࡓࠋ ᰿ᅪᅵተࡣ10-3ࠊ㈓ⶶ᰿ࡣ10-4ࡲ࡛ᕼ㔘ࡋࠊᕼ㔘ᾮࢆࣇࢨ࣒ࣜ࢘⳦㑅ᢥᇵᆅ࡛࠶ࡿ㥖⏣ᇵᆅ ➨3 ⾲ ῧຍࡋ࡚ࠊࣉ࣮ࣞࢺᇵ㣴 25Υ5 ᪥㛫ᬯᡤ ࢆ⾜ࡗࡓࠋᇵ㣴ᚋࠊࢥࣟࢽ࣮ᩘࢆィ ࡋ࡚᰿ᅪ ᅵተཬࡧ㈓ⶶ᰿⤌⧊ෆ࠾ࡅࡿࣇࢨ࣒ࣜ࢘⳦㔞ࢆ௨ୗࡢᘧࡼࡾ⟬ฟࡋࡓࠋ ᰿ᅪᅵተ࣭㈓ⶶ᰿1g ࠶ࡓࡾࡢ⳦㔞 ィ ࢥࣟࢽ࣮ᩘ 10x x:ᕼ㔘ಸ⋡ 6. ᢠ㓟ᶵ⬟ศᯒ (1)ศᯒヨᩱࡢస〇 AMF ᥋✀ 8 㐌㛫ᚋ ᆅୖ㒊ᆅୗ㒊 ཬࡧཎ⳦᥋✀ 8 㐌㛫ᚋ ᆅୗ㒊 ࠊࢧࣥࣉࣜࣥࢢࡋࡓ᳜ ≀యࢆࡑࢀࡒࢀⱼࠊ㈓ⶶ᰿㢮ูࡋࠊᾮయ❅⣲࡛⤖ࡋࡓࠋ (2)SOD άᛶ SOD άᛶࡢ ᐃࡣ➨ 2 ⠇ࡢ 2-2 ࡢ 2 ྠᵝࡢ᪉ἲ࡛⾜ࡗࡓࠋ (3) APX άᛶ APX άᛶࡢ ᐃࡣࠊWu ࡽ㸦2006㸧ࡢ᪉ἲ‽ࡌ࡚⾜ࡗࡓ㸦➨ 12 ᅗ㸧ࠋ⤖ヨᩱ㸦0.1g㸧 50mM ࣜࣥ㓟⦆⾪ᾮ㸦pH7.0㸧3.0ml ࢆῧຍࡋࠊ෭༷ࡋࡓங㖊ࢆ⏝࠸࡚☻○ᚋࠊ㧗㏿㐲ᚰ㸦13,000rpm㸪5Υ㸪 5 ศ㸧ࡋࡓୖΎࢆ⢒㓝⣲ᾮࡋࡓࠋ⢒㓝⣲ᾮ 0.2ml ࣜࣥ㓟⦆⾪ᾮ㸦pH7.0㸧3.0ml ࢆΰྜࡋࠊ10mM ࢫࢥࣝࣅࣥ㓟0.2ml ࢆຍ࠼࡚ 290nm ࠾ࡅࡿ྾ගᗘࢆ ᐃࡋࡓࠋࡑࡢᚋ 10mM H2O2 0.2ml ࢆຍ
27
࠼ࠊ1 ศᚋࡧ 290nm ࠾ࡅࡿ྾ගᗘࢆ ᐃࡋࡓࠋAPX άᛶࡢ⟬ฟ᪉ἲࡣ㸪Nakano and Asada
㸦1981㸧ࡢ᪉ἲᚑ࠸ࠊࢫࢥࣝࣅࣥ㓟ࡢศᏊ྾ගಀᩘ 2.8mM/cm ࢆ⏝࠸࡚㑏ඖᆺࢫࢥࣝࣅࣥ 㓟ྵ㔞ࢆ⟬ฟࡋࠊ1 ศ㛫 APX ࡼࡾ㓟ࡉࢀࡓࢫࢥࣝࣅࣥ㓟㔞࡛⾲ࡋࡓࠋ (4) DPPH ࣛࢪ࢝ࣝᤕᤊ⬟ DPPH ࣛࢪ࢝ࣝᤕᤊ⬟ࡢ ᐃࡣ➨ 2 ⠇ࡢ 2-2 ࡢ 3 ྠᵝࡢ᪉ἲ࡛⾜ࡗࡓࠋ (5) SOD ࢯࢨ࣒ゎᯒ
SOD ࢯࢨ࣒ゎᯒࡣࠊDavis (1964)ཬࡧ Sahoo ࡽ(2007) ࡢ᪉ἲ‽ࡌࠊNative PAGE ࣏
ࣜࢡ࣑ࣜࣝࢻࢤࣝ㟁ẼὋື ࡼࡾ⾜ࡗࡓ㸦➨13 ᅗ㸧ࠋ⏕యヨᩱ 0.5g Extraction buffer (PVPP
75mg/l, Tris 12.1g/l, sucrose 68g/l, EDTA 170mg/l, Tween80 0.031ml/l, ࢳ࢜ࢢࣜࢥ࣮ࣝ㓟ࢼࢺ࣒ࣜ࢘ 800mg/l, pH7.8) ࢆ 3ml ຍ࠼ࠊ෭༷ࡋࡓங㖊࡛ᦶ○ࡋࡓᚋࠊᢳฟᾮࢆ㐲ᚰศ㞳(13,000rpm, 5Υ, 15
ศ)ࡋࡓࠋࡑࡢᚋࠊୖΎ 18μl 40%ࢫࢡ࣮ࣟࢫ 2μl, BPB ⁐ᾮ ࣈࣟࣔࢳ࣮ࣔࣝࣈ࣮ࣝ10mgࢢ
ࣜࢭࣜࣥ1ml␃Ỉ 9ml) 1μl ࢆຍ࠼ࡓࡶࡢࢆヨᩱᾮࡋࡓࠋὋືᵴ AE-6500,ࢺ࣮ᰴᘧ♫
Tris Glycine buffer (Tris 3.0g/l,ࢢࣜࢩࣥ 14.4g/l) ࢆὀࡂࠊ12.5%࣏ࣜࢡ࣑ࣜࣝࢻࢤࣝ
㸦e-PAGEL E-T 12.5L, ࢺ࣮ᰴᘧ♫㸧ࢆタ⨨ࡋࠊヨᩱᾮ 20μl ࢆ࢙࢘ࣝὀධࡋࠊ㟁ẼὋື 㟁
ᅽ:100V, 㟁ὶ:20mA, 130 ศ ࢆ⾜ࡗࡓࠋὋື⤊ᚋࠊBeauchamp and Fridvich (1971)ࡢ᪉ἲ‽
ࡌ࡚ࢤࣝࢆᰁⰍᾮA (Tris 1.97g, NBT 50mg, 1.0M ሷ㓟80ml,␃Ỉ170ml, pH 7.5㹼8)ᾐₕࡋࠊ
ᬯᡤ࡛30 ศ㛫ࠊ࠺ࡋ࡞ࡀࡽᛂࡉࡏࡓࠋᛂ⤊ᚋࠊࢤࣝࢆỈὙࡋࠊᰁⰍᾮ B (Tris 1.97g, 1.0M
ሷ㓟80mlࣜ࣎ࣇࣛࣅࣥ 0.15mg, EDTA 5mg, TEMED 25μl,␃Ỉ 170ml, pH7.5㹼8)ᾐࡋࠊ༑
ศᰁⰍࡉࢀࡿࡲ࡛࠺ࡋࠊᛂ⤊ᚋࠊᙳࢆ⾜ࡗࡓࠋྛ SOD ࢯࢨ࣒ࡢ┦ᑐ㊥㞳 Rf
ࡢホ౯ࡣࠊManganris and Alston (1992)ࡢ᪉ἲ‽ࡌࠊࣂࣥࢻࡢὋື㛤ጞⅬࡽ BPB ⁐ᾮࡢ⛣ື
ᆅⅬࡲ࡛ࡢ㊥㞳ࢆ1 ࡋࠊጞⅬࡽྛࣂࣥࢻࡲ࡛ࡢ┦ᑐ㊥㞳ࢆ⟬ฟࡋࡓࠋ
Fig. 6. Split root system method.
Fig. 7. Treatments of split root system.
Control AMF NPFO
NaCl
AMF㸩NPFO AMF㸩NaCl
L R
L R
L R
AMF NPFOL R
L R
L R
NaCl AMF NPFO AMF NaCl 29Table 3. Composition of Komada’s medium.
Media
Chemicals/compounds
Quantities
(g/l)
Remarks
Komada’s
medium
K
2HPO
41.0
KCl
0.5
Pentachloronitrobenzen,
Na
2B
4O
7࣭
10H
2O, cholic
acid sodium salt and
streptomycin
sulphate
was added finally when
the
medium
was
autoclaved in 1.0 lit, of
distilled water and
cooled. Finally pH has
adjusted to 3.8s0.2 with
10% H
3PO
4.
MgSO
4࣭7H
2O
0.5
Fe-Na-EDTA
0.01
L-asparagine
2.0
D-galactose
20.0
Agar
15.0
Pentachloronitrobenzene
1.0
Na
2B
4O
7࣭10H
2O
10.0
Cholic acid sodium salt
0.5
Streptomycin sulphate
0.3
Test tube for
extracted
enzyme
Test tube for blank (absence
of extracted enzyme)
2.3 ml of 50 mM sodium carbonate buffer
0.1 ml of 1.0 mM NBT solution
0.1 ml of 4.0 mM xanthine solution
0.1 ml of 3.0 mM EDTA solution
0.1 ml of 0.15% BSA solution (100-fold dilution)
0.1 ml of xanthine oxidase solution
0.1 ml of extracted enzyme
0.1 ml of distilled water
30 minutes keep at room temperature in dark condition
0.2 ml of 14 mM copper chloride solution
Measure absorbance of the reaction mixture at 560 nm wave length
Fig. 8. Flow diagram of the procedures in SOD analysis.
Test tube
Test tube (Blank)
0.9 ml of DPPH solution
0.9 ml of MES buffer
0.9 ml of 20% ethanol
0.75 ml of 80% ethanol
0.15 ml of extracted sample
0.15 ml of 80% ethanol
30 minutes keep at 30
oC in dark condition
Measure absorbance of the reaction mixture at 520 nm wave length
Fig. 9. Flow diagram of the procedures in DPPH radical scavenging activity.
Test tube
Test tube (Blank)
0.5 ml of extracted sample
0.5 ml of 0.03% DCIP solution
0.5 ml of 2% thiourea-5% metaphosphoric acid solution
0.25 ml of 2% DNP solution
Keep in water-bath at 50
oC for 70 minutes
1.25 ml of 85% sulfuric acid solution
0.25 ml of 2% DNP solution
30 minutes keep at 30
oC in dark condition
Measure absorbance of the reaction mixture at 520 nm wave length
Fig. 10. Flow diagram of the procedures in ascorbic acid contents assay.
Test tube
Test tube (blank)
3.2 ml of distilled water
0.2 ml of extracted sample
0.2 ml of Folin-Denis solution
0.2 ml of distilled water
0.4 ml of saturated sodium carbonate solution
Keep in dark at 30
oC for 30 minutes
Measure absorbance of the reaction mixture at 700 nm wave length
Fig. 11. Flow diagram of the procedures in polypehnol contents assay.
Test tube
3 ml of 50 mM Phosphate buffer
0.2 ml of extracted enzyme solution
0.2 ml of 10mM ascorbic acid solution
Measure absorbance of the reaction mixture at 290 nm wave length
0.2 ml of 10 mM Hydrogen peroxide solution and keep 1 minutes in
the room temperature
Measure absorbance of the reaction mixture at 290 nm wave length
Fig. 12. Flow diagram of the procedures in APX analysis.
0.5g of fresh
sample
Extraction buffer (pH 7.8) containing:
75 mg/L PVPP
12.1 g/ L Tris
68 g/L sucrose
170 mg /L EDTA
0.031 mL/L Tween 80
800 mg/ L sodium-thioglycolate.
Centrifuge the homogenate at 15 min in 5°C and use the supernatant
for electrophoresis
Run at 100V, 20mM and 130 min
Mix extracted enzyme, 18 μl : sucrose, 2 μl : BPB, 1 μl. From there
apply 20 μl for each well
Staining A (pH 7.5 to 8)
Tris, 1.97 g + NBT, 50 mg + HCl, 80 ml + D. water, 170 ml
Duration 30 minutes in dark at room temperature then washed by
D.water once. Use shaker during the time of staining.
Staining B (pH 7.5 to 8)
Tris, 1.97 g + HCl, 80 ml + D. water, 170 ml + ribo flavin, 1 mg +
EDTA, 5 mg + TEMED, 25 μl
Use shaker and 15W two florescent lamps during the time of staining.
Stain until bands are visualized.
Fig. 13. Flow diagram of the procedures in SOD isozyme analysis.
37 ⤖ ᯝ ➨1 ⠇ ࢫࣃࣛ࢞ࢫ࡛ࡢ AMF ࡼࡿ❧ᯤ⪏ᛶㄏᑟࡢရ✀࣭⳦✀㛫ᕪ AMF ᥋✀ 12 㐌㛫ᚋࡢᆅୖ㒊ࠊᆅୗ㒊≀㔜ࡶࠊAMF ฎ⌮༊࡛ᑐ↷༊ࡼࡾቑࡋ࡚࠸ࡓ㸦➨ 14,15 ᅗ㸧ࠋ≉͂ࢫ࣮ࣃ࣮࢙࣒࢘ࣝ࢝̓ࠊ͂㞝࢞ࣜࣂ࣮̓ࠊ͂‶⣸̓ࡢ AMF ฎ⌮༊࡛ࡣᆅ ୖ㒊࣭ᆅୗ㒊ࡶᑐ↷༊ࡼࡾࡁࡃቑຍࡋ࡚࠸ࡓࠋ❧ᯤ᥋✀6 㐌㛫ᚋࡢⓎㄪᰝࡢ⤖ᯝࠊ ရ✀ࡢAMF ᥋✀༊࡛ᑐ↷༊ࡼࡾࡶⓎ⛬ᗘࡀ㍍ῶࡉࢀࡓ㸦➨ 16 ᅗ㸧ࠋ≉࢙࣒͂࢘ࣝ࢝̓ࠊ͂ࢫ ࣮ࣃ࣮࢙࣒࢘ࣝ࢝̓ࠊ͂‶⣸̓ࡢⓎᗘࡣ 20 ௨ୗప࠸್࡛࠶ࡾࠊᑐ↷༊ࡢⓎᗘࡢ 50%⛬ ᗘ࡛࠶ࡗࡓࠋ❧ᯤ⳦᥋✀ 6 㐌㛫ᚋࡢ AMF ᥋✀༊ࡢᆅୖ㒊≀㔜ࡣࠊቑຍࡍࡿሙྜࡀከࡃࡳࡽ ࢀࡓ㸦➨17,18 ᅗ㸧ࠋࡲࡓࠊᆅୗ㒊≀㔜ࡣ࡚ࡢရ✀࠾࠸࡚ AMF ༊࡛ᑐ↷༊ࢆୖᅇࡗࡓࠋࡇ ࡢሙྜࠊ≉࢙࣒͂࢘ࣝ࢝̓ࡢᆅୖ㒊ࠊ͂㞝࢞ࣜࣂ࣮̓͂ࣂ࣮࢞ࣥࢹ࣮̓ࡢᆅୗ㒊≀㔜 ࡛ࡣ㢧ⴭቑࡋ࡚࠸ࡓࠋ
➨2 ⠇ Split root system ἲࡼࡿࢫࣃࣛ࢞ࢫ࡛ࡢ⏕≀࣭Ꮫⓗᡭἲࡼࡿㄏᑟᢠᛶホ౯ཬࡧ
ᢠ㓟 ᶵ⬟ኚືゎᯒ 1㸬᳜≀య⏕⫱ཬࡧ AMF ឤᰁ⋡ ❧ᯤ⳦᥋✀๓࠾࠸࡚ࠊᆅୖ㒊ཬࡧᆅୗ㒊≀㔜ࡣ≉AMF ༊ཬࡧ NPFO ༊࡛ᑐ↷༊ࡼࡾ ቑࡋࡓ㸦➨19 ᅗ㸧ࠋ❧ᯤ⳦᥋✀ᚋ࠾࠸࡚ࠊᆅୗ㒊≀㔜ࡣྛฎ⌮༊࡛ᑐ↷༊ࡼࡾቑࡋࡓ ሙྜࡀከࡗࡓࠊ≉AMF ༊ ➨ 27 ᅗ ࡇࡢሙྜࠊᆅୗ㒊≀㔜ࡘ࠸࡚ࡣࠊฎ⌮᰿㸦R㸧ཬࡧ ↓ฎ⌮᰿㸦L㸧ࡶᑐ↷༊ࢆୖᅇࡗࡓࠋ୍᪉ࠊNaCl ฎ⌮ࡼࡿ AMF ឤᰁ⋡ࡢኚࡣࡳࡽࢀ࡞ ࡗࡓࠋ 2㸬❧ᯤⓎホ౯ ᑐ↷༊࡛ࡣ࡚ࡢಶయࡀⓎࡋ࡚࠾ࡾࠊ⨯ᣦᩘࡀ70 ௨ୖ࡞ࡗࡓࠋࡑࢀᑐࡋࠊAMFࠊ㠀 ཎᛶࣇࢨ࣒ࣜ࢘⳦ཬࡧ NaCl ࡢྛฎ⌮༊࠾࠸࡚ࠊⓎ⋡Ⓨ⛬ᗘࡢ㍍ῶࡀࡳࡽࢀࡓ㸦➨
38 24,25,26 ᅗ㸧ࠋࡇࡢሙྜࠊ≉ AMF ༊࠾࠸࡚ࠊฎ⌮᰿㸦R㸧ཬࡧ↓ฎ⌮᰿㸦L㸧ࡶⓎ⛬ᗘ ࡀ㢧ⴭపୗࡋࠊ⨯ᣦᩘࡀ40 ௨ୗ࡞ࡗࡓࠋNPFO ༊࠾࠸࡚ࡣࠊฎ⌮᰿㸦R㸧ཬࡧ↓ฎ⌮᰿ 㸦L㸧ࡶⓎ⛬ᗘࡶపୗࡋ⨯ᣦᩘࡀ 50 ௨ୗ࡞ࡗࡓࠋNaCl ༊࠾࠸࡚ࡣࠊฎ⌮᰿㸦R㸧࠾ ཬࡧ↓ฎ⌮᰿㸦L㸧ࡶⓎ⛬ᗘࡶపୗࡋࠊ⨯ᣦᩘࡀ 50 ௨ୗ࡞ࡗࡓࠋAMF-NPFO ే⏝༊ ࠾࠸࡚ࠊฎ⌮᰿㸦R㸧ཬࡧ↓ฎ⌮᰿㸦L㸧ࡶⓎ⛬ᗘࡢ㍍ῶࡀࡳࡽࢀࠊⓎ⋡ࡀࡁࡃపୗࡍ ࡿሙྜࡀ࠶ࡗࡓࠋࡲࡓࠊAMF-NaCl ే⏝༊࠾࠸࡚ࡶࠊฎ⌮᰿㸦R㸧ཬࡧ↓ฎ⌮᰿㸦L㸧ࡶⓎ ⛬ᗘࡢ㍍ῶࡶࡳࡽࢀࠊⓎ⋡ࡀࡁࡃపୗࡍࡿሙྜࡀ࠶ࡗࡓࠋ 3㸬ᢠ㓟ᶵ⬟ኚືゎᯒ (1) SOD άᛶ SOD άᛶࡘ࠸࡚ࡣࠊ❧ᯤ⳦᥋✀๓࠾࠸࡚ࠊᆅୖ㒊ཬࡧᆅୗ㒊㸦LࠊR㸧 ࡢ SOD άᛶࡣ ᑐ↷༊ࡼࡾAMFࠊNPFOࠊNaCl ࡼࡗ࡚㧗ࡲࡿሙྜࡀ࠶ࡗࡓ㸦➨ 20 ᅗ㸧ࠋࡇࡢሙྜࠊ≉ AMF
༊ཬࡧNPFO ༊࠾ࡅࡿ SOD άᛶࡢቑࡀ㢧ⴭ࡛࠶ࡗࡓࠋ❧ᯤ⳦᥋✀ᚋ࠾࠸࡚ࡣࠊAMFࠊ
NPFOࠊNaCl ࡼࡿᆅୗ㒊ࡢ SOD άᛶࡣฎ⌮᰿㸦R㸧ཬࡧ↓ฎ⌮᰿㸦L㸧ࡢ୧᪉࡛ᑐ↷༊ࡼࡾ ࡁࡃቑຍࡋࡓ㸦➨28 ᅗ㸧ࠋ (2) DPPH ࣛࢪ࢝ࣝᤕᤊ⬟ DPPH ࣛࢪ࢝ࣝᤕᤊ⬟ࡘ࠸࡚ࡣࠊ❧ᯤ⳦᥋✀๓࠾࠸࡚ࠊAMF-NaCl ే⏝༊ࡢᆅୖ㒊࠾ ࠸࡚ࡣኚࡀࡳࡽࢀࡎࠊAMFࠊNPFOࠊNaCl ཬࡧ AMF-NPFO ే⏝ࡼࡗ࡚ᆅୖ㒊ཬࡧᆅୗ㒊ฎ ⌮᰿(R)ཬࡧ↓ฎ⌮᰿(L)ࡢ୧᪉࡛ᑐ↷༊ࡼࡾቑຍࡋࡓ㸦➨ 21 ᅗ㸧ࠋ❧ᯤ⳦᥋✀ᚋ࠾࠸࡚ࡣࠊ ᆅୗ㒊ࡢฎ⌮᰿(R)ཬࡧ↓ฎ⌮᰿(L)ࡢ୧᪉࡛ቑຍഴྥࡀࡳࡽࢀࡓ㸦➨ 29 ᅗ㸧ࠋ (3) ⥲ࢫࢥࣝࣅࣥ㓟ྵ㔞 ⥲ࢫࢥࣝࣅࣥ㓟ྵ㔞ࡘ࠸࡚ࡣࠊ❧ᯤ⳦᥋✀๓࠾࠸࡚ࠊᆅୖ㒊ཬࡧᆅୗ㒊㸦LࠊR㸧 ࡶࠊAMFࠊNPFOࠊNaCl ࡼࡗ࡚ᑐ↷༊ࡼࡾ㧗ࡲࡿሙྜࡀ࠶ࡗࡓ㸦➨ 22 ᅗ㸧ࠋࡇࡢሙྜࠊ≉
39
AMF ༊ཬࡧ NaCl ༊࠾࠸࡚ቑࡀ㢧ⴭ࡛࠶ࡗࡓࠋ❧ᯤ⳦᥋✀ᚋ࠾࠸࡚ࡣࠊAMFࠊNPFOࠊ NaCl ࡼࡗ࡚ᆅୗ㒊ࡢฎ⌮᰿㸦R㸧ཬࡧ↓ฎ⌮᰿㸦L㸧ࡢ୧᪉࡛ᑐ↷༊ࡼࡾࡁࡃቑຍࡋࡓ㸦➨ 30 ᅗ㸧ࠋ (4) ⥲࣏ࣜࣇ࢙ࣀ࣮ࣝྵ㔞 ⥲࣏ࣜࣇ࢙ࣀ࣮ࣝྵ㔞ࡘ࠸࡚ࡣࠊ❧ᯤ⳦᥋✀๓࠾࠸࡚ࠊAMFࠊNPFOࠊNaCl ࡼࡿᆅୖ 㒊ࡢቑࡀ㢧ⴭ࡛࠶ࡗࡓ㸦➨23 ᅗ㸧ࠋᆅୗ㒊࠾࠸࡚ࡣࠊฎ⌮᰿㸦R㸧ཬࡧ↓ฎ⌮᰿㸦L㸧ࡢ୧᪉ ࡛㧗ࡲࡿሙྜࡀ࠶ࡾࠊ≉AMF ༊ཬࡧ AMF-NaCl ే⏝༊࡛ࡣࡁࡃቑຍࡋࡓࠋ❧ᯤ⳦᥋✀ᚋ ࠾࠸࡚ࡣࠊྛฎ⌮༊࡛ᆅୗ㒊ࡢฎ⌮᰿㸦R㸧ཬࡧ↓ฎ⌮᰿㸦L㸧ࡢࡕࡽ࠾࠸࡚ࡶᑡࡋቑຍࡋ ࡓ㸦➨31 ᅗ㸧ࠋ ➨3 ⠇ AMF ඹ⏕ࢫࣃࣛ࢞ࢫ࠾ࡅࡿ❧ᯤ⪏ᛶㄏᑟඹ⏕≉␗ⓗ SOD ࢯࢨ࣒ኚື 1㸬AMF ࡼࡿ❧ᯤ⪏ᛶホ౯ ⏕⫱ㄪᰝࡢ⤖ᯝࠊ❧ᯤ⳦᥋✀๓ᚋ࠾࠸࡚ࠊᆅୖ㒊ཬࡧᆅୗ㒊≀㔜ࡣ AMF ༊ࡀᑐ↷༊ࢆ ᭷ពୖᅇࡾࠊ≉Gi ༊ࡢቑࡀ㢧ⴭ࡛࠶ࡗࡓ㸦➨ 32,33,39 ᅗ㸧ࠋ❧ᯤ⳦᥋✀ 8 㐌㛫ᚋࡢⓎ ホ౯ࡢ⤖ᯝࠊGi ༊ཬࡧ GM ༊ࡶᑐ↷༊ẚ࡚ AMF ༊࡛Ⓨ⋡Ⓨ⛬ᗘࡢ㍍ῶࡀࡳࡽࢀ ࡓ㸦➨37,38 ᅗ㸧ࠋࡇࡢሙྜࠊ≉ Gi ༊࠾࠸࡚Ⓨ⛬ᗘࡀ㢧ⴭపୗࡋࠊ⨯ᣦᩘࡀ 10 ௨ୗ ࡞ࡾࠋGM ༊࠾࠸࡚ࡣ⨯ᣦᩘࡀ 20 ௨ୗ࡞ࡗࡓࠋ 2㸬⳦㔞ࡢㄪᰝ ᰿ᅪᅵተཬࡧ᰿⤌⧊ෆ࠾ࡅࡿ❧ᯤ⳦㔞ࢆㄪᰝࡋࡓ⤖ᯝࠊ᰿ᅪᅵተ࡛ࡣ Gi ༊ཬࡧ GM ༊ ࠾࠸࡚⳦㔞ῶᑡࡀࡳࡽࢀࡓࠋࡇࡢሙྜࠊ≉GM ༊࡛⳦㔞ῶᑡࡀ㢧ⴭࡳࡽࢀࡓ ➨ 4 ⾲ ࠋ୍᪉ࠊ ᰿⤌⧊ෆ࡛ࡶGi ༊ཬࡧ GM ༊࠾࠸࡚⳦㔞ῶᑡࡀࡳࡽࢀࠊ≉ GM ༊ࡢ⳦㔞ῶᑡࡀ㢧ⴭ࡛࠶ࡗ ࡓࠋ
40 3㸬SOD άᛶ SOD άᛶࡘ࠸࡚ࡣࠊ❧ᯤ⳦᥋✀๓࠾࠸࡚ࠊAMF ༊࡛ࡣᆅୖ㒊࡛ᑐ↷༊ࡼࡾࡶ᭷ពቑ ࡋࠊᆅୗ㒊࠾࠸࡚ࡶቑຍഴྥ࠶ࡾࠊ≉Gi ༊ࡢቑຍࡀ㢧ⴭ࡛࠶ࡗࡓ㸦➨ 34 ᅗ㸧ࠋ❧ᯤ⳦ ᥋✀ᚋ࠾࠸࡚ࡣࠊᆅୗ㒊ࡢSOD άᛶࡀ Gi ༊࠾࠸࡚᭷ពቑࡋࠊGM ༊࠾࠸࡚ࡶቑຍഴ ྥ࠶ࡗࡓ㸦➨40 ᅗ㸧ࠋ 4㸬APX άᛶ APX άᛶࡘ࠸࡚ࡣࠊ❧ᯤ⳦᥋✀๓࠾࠸࡚ࠊAMF ༊࡛ࡣᆅୖ㒊࡛ࡣ᭷ពቑࡋࠊᆅୗ 㒊࠾࠸࡚ࡶቑຍഴྥ࠶ࡾࠊ≉Gi ༊ࡢቑຍࡀ㢧ⴭ࡛࠶ࡗࡓ㸦➨ 35 ᅗ㸧ࠋ❧ᯤ⳦᥋✀ᚋ࠾ ࠸࡚ࡣࠊᆅୗ㒊ࡢAPX άᛶࡀ Gi ༊࠾࠸࡚᭷ពቑࡋࠊGM ༊࠾࠸࡚ࡶቑຍഴྥ࠶ࡗࡓ 㸦➨41 ᅗ㸧ࠋ 5㸬DPPH ࣛࢪ࢝ࣝᤕᤊ⬟ DPPH ࣛࢪ࢝ࣝᤕᤊ⬟ࡘ࠸࡚ࡣࠊ❧ᯤ⳦᥋✀๓࠾࠸࡚ࠊAMF ༊ࡢᆅୖ㒊ཬࡧᆅୗ㒊࠾ ࠸࡚ቑຍഴྥࡀࡳࡽࢀࡓࠊࡇࡢሙྜࠊ≉GM ༊ࡢቑࡀ㢧ⴭ࡛࠶ࡗࡓ㸦➨ 36 ᅗ㸧ࠋ❧ᯤ⳦᥋ ✀ᚋ࠾࠸࡚ࡣࠊGi ༊ཬࡧ GM ༊࠾࠸࡚ᆅୗ㒊ࡢ DPPH ࣛࢪ࢝ࣝᤕᤊ⬟ቑຍഴྥࡀࡳࡽࢀࡓ 㸦➨42 ᅗ㸧ࠋ 6. SOD ࢯࢨ࣒ゎᯒ
㈓ⶶ᰿࠾ࡅࡿSOD ࢯࢨ࣒ゎᯒࢆ⾜ࡗࡓ⤖ᯝࠊCu/ZnSODs ཬࡧ Mn/Fe-SODs ࡀ❧ᯤ
⳦᥋✀๓ᚋ࠾࠸࡚AMF ༊ཬࡧᑐ↷༊࡛ࡶ᳨ฟࡉࢀࡓ㸦➨ 43 ᅗ㸧ࠋࡇࡢሙྜࠊࡍ࡚ࡢฎ⌮
༊࠾࠸࡚Mn/Fe ࡼࡾ Cu/ZnSOD ࣂࣥࢻࣃࢱ࣮ࣥࡀᙉࡃࡳࡽࢀࠊࣂࣥࢻࣃࢱ࣮ࣥࡢ⨨ࡘ࠸࡚
ࡣฎ⌮༊㛫ࡢᕪࡀࡳࡽࢀ࡞ࡗࡓࠋ❧ᯤ⳦᥋✀๓㸦Foa-㸧࠾࠸࡚ࠊSOD-1 㸦Rf = 0.37, Cu/Zn-SOD㸧
ࡣGi ཬࡧ GM ༊࡛ᑐ↷༊ẚ࡚ᙉ࠸Ⓨ⌧ࡀࡳࡽࢀࡓࠋ❧ᯤ⳦᥋✀ 8 㐌㛫ᚋ㸦Foa+㸧࠾࠸
41 ࡀࡳࡽࢀࠊᑐ↷༊࡛ࡣ❧ᯤ⳦᥋✀๓ࡼࡾCu/Zn ࣂࣥࢻࣃࢱ࣮ࣥࡀᙅࡃ࡞ࡗࡓሙྜࡀࡳࡽࢀࡓࠋ ࡲࡓࠊSOD-3 (Rf = 0.25, Mn/Fe-SOD)ࡣ GM ༊࡛ᑐ↷༊ẚ࡚ᙉ࠸Ⓨ⌧ࡀࡳࡽࢀࡓࠋ
Fig. 14. Growth enhancement in mycorrhizal (Glomus intraradices, Gi) asparagus plants.
Control Gi
‘Manmi murasaki’
‘Super welcome’
Control Gi
42‘Welcome’ ‘Super Welcome’
‘Zenyu
Guliber’ ‘Burgundy’ ‘Manmi murasaki’
0
0.1
0.2
0.3
0.4
0
0.2
D
ry
w
eight
of
shoots
(g)
D
ry
w
eight of
roots (g)
Fig. 15. Dry weight of shoots and roots in asparagus plants before Fusarium
oxysporum f. sp. asparagi (MAFF3055567) inoculation. , control; , Glomus
intraradices. Bars represent standard errors (n=5). Columns denoted by different
letters indicate significant difference according to Tukey's test (P<0.05).
0.4
de
c
d
a
e
b
b
c
e
b
ab
ab
c
a
cd
b
a
ab
c
b
430
20
40
60
80
100
0
20
40
60
80
Incidence of
Fusarium
root
rot
Fig. 16. Disease incidece and index of Fusarium root rot in asparagus plants 6 weeks after
Fusarium oxysporum f. sp. asparagi (MAFF 305556) inoculation. Ratio of diseased storage
roots: , ~20; , 20~40; , 40~60 ; , 60~80; , 80~100 (%). , Control; , Glomus
intraradices. Bars represent standard errors (n=5). Columns denoted by different letters indicate
significant difference according to Tukey's test (P<0.05).
C Gi ‘Welcome’ ‘Super Welcome’ ‘Zenyu Guliber’ ‘Burgundy’ ‘Manmi murasaki’
D
isease index
of Fusarium
root
rot
a
ab
b
b
c
d
d
e
ef
f
C Gi C Gi C Gi C Gi ‘Welcome’ ‘Super Welcome’ ‘Zenyu Guliber’ ‘Burgundy’ ‘Manmi murasaki’ 44Fig. 17 Asparagus plants (‘Welcome’) after Fusarium oxysporum f. sp. asparagi
(MAFF 305556) inoculation.
Control Glomus intraradices
‘Welcome’ ‘Super Welcome’ ‘Zenyu Guliber’ ‘Burgundy’ ‘Manmi murasaki’
0
0.1
0.2
0.3
0.4
0.5
0
0.2
0.4
0.6
0.8
D
ry
w
eight
of
shoots
(g)
Dry
weight
of
roots (g)
Fig. 18. Dry weight of shoots and roots in asparagus plants after Fusarium
oxysporum f. sp. asparagi (MAFF 305556) inoculation. , control; , Glomus
intraradices. Bars represent standard errors (n=5). Columns denoted by different
letters indicate significant difference according to Tukey's test (P<0.05).
a
f
f f
d
c c
b
d
e
d
bc
d
cd
cd
a a
c
b
bc
460
0.5
1
1.5
2
0
0.2
0.4
0.6
0.8
C AMF NPFO AMF+NPFO NaCl AMF+NaCl
D
ry
w
eight
of
shoots
(g)
D
ry
w
eight of
roots (g)
Fig. 19. Dry weight of shoots and roots of asparagus plants before Fusarium
oxysporum f. sp. asparagi inoculation. C, control; AMF, Glomus intraradices;
NPFO, Non-pathogenic Fusarium oxysporum f. sp. asparagi; NaCl, 50 mM
NaCl. , non-treated root parts with㻌 AMF, NPFO and NaCl; , treated root
parts with㻌 AMF, NPFO and NaCl.㻌 Bars represent standard errors (n =10).
Columns denoted by different letters indicate significant difference according
to Tukey's test (P<0.05).
b
b b
a
ab
a
b
a
a
c bca
b
a
a
ab
a
b
ab
b
470
50
100
150
200
0 50 100 150 200 250 0 50 100 150 2000
50
100
150
200
C AMFSO
D
activit
y in shoots
(units/
g FW
)
b a b b a a 0 100 200 300 C NPFO AMF+NPFO 0 50 100 150 200 C NaCl AMF+NaClSO
D
activit
y in roots
(units/
g FW
)
b
a
c
b
b
a
a
a
a
b
ab
a
a a
ab
b
a
a
Fig. 20. Superoxide dismutase (SOD) activity in shoots and roots before Fusarium
oxysporum f. sp. asparagi inoculation. C, control; AMF, Glomus intraradices; NPFO,
Non-pathogenic Fusarium oxysporum f. sp. asparagi; NaCl, 50 mM NaCl. ,
non-treated root parts; , non-treated root parts. Bars represent standard errors (n=10). Columns
denoted by different letters indicate significant difference according to Tukey's test
(P<0.05).
DPPH
radi
cal
scavenging
activity
in shoots
(m
g/g FW
)
0 0.02 0.04 0.06 0 0.02 0.04 0.06 0.08 0.1 C AMFb
a
c
c
b
a
Fig. 21. 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity in shoots and roots
before Fusarium oxysporum f. sp. asparagi inoculation. C, control; AMF, Glomus intraradices;
NPFO, Non-pathogenic Fusarium oxysporum f. sp. asparagi; NaCl, 50 mM NaCl. ,
non-treated root parts; , non-treated root parts. Bars represent standard errors (n=10). Columns
denoted by different letters indicate significant difference according to Tukey's test (P<0.05).
DPPH
radi
cal
scavenging
activity
in
roots
(m
g/g
FW
)
0 0.02 0.04 0.06 0 0.02 0.04 0.06 0.08 0.1 C NPFO AMF+NPFOb
a
c
a
b
b
c
d
d
0 0.01 0.02 0.03 0.04 0 0.02 0.04 0.06 0.08 0.1 C NaCl AMF+NaClb
a
ab
b
b
a a
a
a
490 1 2 3 4 0 5 10 15 20 25 C NaCl AMF+NaCl
Fig. 22. Ascorbic acid contents in shoots and roots before Fusarium oxysporum f. sp.
asparagi inoculation. C, control; AMF, Glomus intraradices; NPFO,
Non-pathogenic Fusarium oxysporum f. sp. asparagi; NaCl, 50 mM NaCl. ,
non-treated root parts; , non-treated root parts. Bars represent standard errors (n=10).
Columns denoted by different letters indicate significant difference according to
Tukey's test (P<0.05).
Ascorbic acid
contents
in roots
(m
g/g
FW
)
Asco
rb
ic acid
co
nten
ts
in shoots
(m
g/g F
W
)
0 1 2 3 4 5 0 5 10 15 20 25 C AMFb
a
b b
a
a
0 1 2 3 4 0 5 10 15 20 C NPFO AMF+NPFOb
a
c
b b
a
a
a
a
b
a
c
b b
a
a
ab
a
500 10 20 30 40 0 20 40 60 C AMF
Poly
phenol
contents
in shoots
(m
g/g FW
)
Poly
phenol contents
in roots
(m
g/g
FW
)
Fig. 23. Polyphenol contents in shoots and roots before Fusarium oxysporum f. sp.
asparagi inoculation. C, control; AMF, Glomus intraradices; NPFO,
Non-pathogenic Fusarium oxysporum f. sp. asparagi; NaCl, 50 mM NaCl. ,
non-treated root parts; , non-treated root parts. Bars represent standard errors (n=10).
Columns denoted by different letters indicate significant difference according to
Tukey's test (P<0.05).
b
a
c c
b
a
0 10 20 30 40 0 10 20 30 40 C NPFO AMF+NPFOb
a
c
d d
cd
a
bc
b
0 10 20 30 40 0 15 30 45 C NaCl AMF+NaClb
a
a
c c
b b
b
a
510 20 40 60 80 100 L R L R L R L R L R L R
Incidence of
Fusarium
ro
ot
ro
t (%)
Fig. 24. Incidence of Fusarium root rot in asparagus plants 4 weeks after
Fusarium oxysporum f. sp. asparagi (MAFF305556) inoculation. C, control;
AMF, Glomus intraradices; NPFO, Non-pathogenic Fusarium oxysporum f. sp.
asparagi; NaCl, 50 mM NaCl; L, non-treated root parts with AMF, NPFO and
NaCl; R, treated root parts with AMF, NPFO and NaCl. Ratio of diseased
storage roots; , -20; , 20-40 ; , 40-60; , 60-80; 㻌 , 80-100 ( % ).
C AMF NPFO AMF+NPFO NaCl AMF+NaCl
0 20 40 60 80 100 L R L R L R L R L R L R
Fig. 25. Disease index of Fusarium root rot in asparagus plants 4 weeks after
Fusarium oxysporum f. sp. asparagi (MAFF305556) inoculation. C, control;
AMF, Glomus intraradices; NPFO, Non-pathogenic Fusarium oxysporum f.
sp. asparagi; NaCl, 50 mM NaCl; L, non-treated root parts with AMF,
NPFO and NaCl; R, treated root parts with AMF, NPFO and NaCl. Bars
represent standard errors (n=10). Columns denoted by different letters
indicate significant difference according to Tukey's test (P<0.05).
Disease in
dex
of
F
usarium
root
rot
C AMF NPFO AMF+NPFO NaCl AMF+NaCl
a
a
d
de
b
bc
b
b
c
c
b
e
53Fig. 26. Asparagus roots after Fusarium oxysporum f. sp. asparagi inoculation.
Control༊༊ Control༊
AMF༊ AMF༊
AMF+ AMF+ AMF䠉
AMF䠉
0 0.2 0.4 0.6 0.8
C AMF NPFO AMF+NPFO NaCl AMF+NaCl
