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   <front>
      <journal-meta>
         <journal-id journal-id-type="publisher-id">pnut</journal-id>
         <journal-title-group>
            <journal-title>Peanut Science</journal-title>
         </journal-title-group>
         <issn pub-type="ppub">0095-3679</issn>
         <issn pub-type="active">0095-3679</issn>
         <publisher>
            <publisher-name>American Peanut Research and Education Society</publisher-name>
            <publisher-loc/>
         </publisher>
      </journal-meta>
      <article-meta>
         <article-id pub-id-type="doi">10.3146/0095-3679-43.1.49</article-id>
         <article-id pub-id-type="publisher-id">0095-3679-43.1.49</article-id>
         <article-categories>
            <subj-group subj-group-type="heading">
               <subject>ARTICLES</subject>
            </subj-group>
         </article-categories>
         <title-group>
            <article-title>Evidence for a Second RKN Resistance Gene in Peanut</article-title>
            <alt-title alt-title-type="running-head">S<sc>econd</sc> RKN R<sc>esistance</sc> G<sc>ene</sc>
            </alt-title>
         </title-group>
         <contrib-group>
            <contrib contrib-type="author" xlink:type="simple">
               <name name-style="western">
                  <surname>Branch</surname>
                  <given-names>W.D.</given-names>
                  <x xml:space="preserve"> </x>
               </name>
               <xref ref-type="aff" rid="aff1">
                  <sup>1</sup>
               </xref>
               <xref ref-type="corresp" rid="cor1">
                  <sup>*</sup>
               </xref>
               <x xml:space="preserve">, </x>
            </contrib>
            <contrib contrib-type="author" xlink:type="simple">
               <name name-style="western">
                  <surname>Brenneman</surname>
                  <given-names>T.B.</given-names>
                  <x xml:space="preserve"> </x>
               </name>
               <xref ref-type="aff" rid="aff2">
                  <sup>2</sup>
               </xref>
               <x xml:space="preserve">, and </x>
            </contrib>
            <contrib contrib-type="author" xlink:type="simple">
               <name name-style="western">
                  <surname>Noe</surname>
                  <given-names>J.P.</given-names>
                  <x xml:space="preserve"> </x>
               </name>
               <xref ref-type="aff" rid="aff3">
                  <sup>3</sup>
               </xref>
            </contrib>
            <aff id="aff1">
					          <label>
                  <sup>1</sup>
               </label>Professor, University of Georgia, Dept. of Crop &amp; Soil Sciences, Coastal Plain Experiment Station, 2360 Rainwater Road, Tifton, GA 31793-5766
				</aff>
            <aff id="aff2">
					          <label>
                  <sup>2</sup>
               </label>Professor, University of Georgia, Dept. of Plant Pathology, Coastal Plain Experiment Station, 2360 Rainwater Road, Tifton, GA 31793-5766
				</aff>
            <aff id="aff3">
					          <label>
                  <sup>3</sup>
               </label>Professor, University of Georgia, Dept. of Plant Pathology, Athens, GA 30602
				</aff>
         </contrib-group>
         
         <pub-date pub-type="ppub">
            <month>April</month>
            <year>2016</year>
         </pub-date>
         <volume>43</volume>
         <issue>1</issue>
         <fpage>49</fpage>
         <lpage>51</lpage>
         <permissions>
            <copyright-statement>© 2016 American Peanut Research and Education Society</copyright-statement>
            <copyright-year>2016</copyright-year>
         </permissions>
         <related-article related-article-type="pdf"
                          xlink:href="0095-3679-43.1.49.pdf"
                          xlink:type="simple"/>
         <abstract>
            <title>ABSTRACT</title>
            <p>Root-knot nematode (RKN), [<italic>Meloidogyne arenaria</italic> (Neal) Chitwood race 1] can result in highly significant yield losses in peanut (<italic>Arachis hypogaea</italic> L.) production. Fortunately, very high levels of RKN nematode resistance have been identified and incorporated from wild species into newly developed peanut cultivars. In 2011-12 at Tifton, GA, a field site was artificially inoculated with <italic>M. arenaria</italic> race 1. A susceptible cultivar was used to uniformly increase the peanut-specific race 1 nematode population during the summer and fall; whereas, hairy vetch (<italic>Vicia villosa</italic> Roth) was used for the same purpose each winter as a susceptible cover crop. During 2013 and 2014, space-planted F<sub>2</sub> and F<sub>3</sub> populations from cross combinations involving <italic>A. hypogaea</italic> susceptible × resistant parental lines derived from ‘COAN’ were evaluated, respectively. Several past inheritance studies had suggested a single dominant gene, R<italic>ma</italic>, controlled the resistance. However in this study, the occurrence of a second recessive gene (<italic>rma<sub>2</sub>
               </italic>) was also found to be involved in very high peanut RKN resistance. Inheritance data fit a 13:3 genetic model and confirmed an earlier report for two RKN-resistance genes (<italic>Rma<sub>1</sub>
               </italic> and <italic>rma<sub>2</sub>
               </italic>) found in TxAG-6 and now COAN.</p>
         </abstract>
         <kwd-group>
            <title>Key Words<x xml:space="preserve">:</x>
            </title>
            <kwd>
               <italic>Arachis hypogaea</italic> L.</kwd>
            <x xml:space="preserve">, </x>
            <kwd>groundnut</kwd>
            <x xml:space="preserve">, </x>
            <kwd>inheritance data</kwd>
            <x xml:space="preserve">, </x>
            <kwd>
               <italic>Meloidogyne arenaria</italic> (Neal) Chitwood race 1</kwd>
            <x xml:space="preserve">, </x>
            <kwd>genetic ratio</kwd>
            <x xml:space="preserve">.</x>
         </kwd-group>
         <counts>
            <fig-count count="0"/>
            <table-count count="3"/>
            <ref-count count="20"/>
            <page-count count="3"/>
         </counts>
      </article-meta>
   </front>
   <body>
      <sec id="s1">
			<title>Introduction</title>
         <p>Root-knot nematodes (RKN) [<italic>Meloidogyne arenaria</italic> (Neal) Chitwood race 1] are a costly peanut (<italic>Arachis hypogaea</italic> L.) production problem in the U.S. (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b14">Kokalis-Burelle and Rodriguez-Kabana 1997</xref>; <xref ref-type="bibr" rid="i0095-3679-43-1-49-b20">Starr <italic>et al.</italic>, 2002</xref>). Introgression of RKN resistance from the wild species resulted in the first improved peanut cultivar ‘COAN’ (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b16">Simpson and Starr 2001</xref>) with a very high RKN resistance. Since then, several other RKN-resistant cultivars have been developed from COAN:‘NemaTAM’ (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b18">Simpson <italic>et al.</italic> 2003</xref>), ‘Webb’ (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b19">Simpson <italic>et al.</italic> 2013</xref>); ‘Tifguard’ (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b13">Holbrook <italic>et al.</italic> 2008</xref>); ‘Georgia-14N’ (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b4">Branch and Brenneman 2015</xref>), and ‘TifNV-High O/L’.</p>
         <p>Inheritance of the very high RKN resistance found in COAN was first reported to be controlled as a single dominant gene in several studies (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b7">Burow <italic>et al.</italic> 1996</xref>; <xref ref-type="bibr" rid="i0095-3679-43-1-49-b8">Choi <italic>et al.</italic> 1999</xref>; <xref ref-type="bibr" rid="i0095-3679-43-1-49-b11">Chu <italic>et al.</italic> 2011</xref>; <xref ref-type="bibr" rid="i0095-3679-43-1-49-b9">Church <italic>et al.</italic> 2000</xref>). <xref ref-type="bibr" rid="i0095-3679-43-1-49-b15">Nagy <italic>et al.</italic> (2010)</xref> proposed the name <italic>Rma</italic> for this single dominant RKN resistance gene.</p>
         <p>In 1996, Garcia <italic>et al</italic>. suggested two dominant genes (<italic>Mae</italic> and <italic>Mag)</italic> controlling egg number and galling, respectively. However, the F<sub>2</sub> data were derived from different parental material than COAN. In yet another genetic study by <xref ref-type="bibr" rid="i0095-3679-43-1-49-b10">Church <italic>et al.</italic> (2005)</xref>, a recessive gene in addition to the single dominant gene was also proposed involving TxAG-6 (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b17">Simpson <italic>et al.</italic>, 1993</xref>), the interspecific hybrid that served as the source of RKN resistance. The objective of current genetic study was to further determine the inheritance of a proposed second RKN resistance gene found in peanut derived from COAN.</p>
      </sec>
      <sec id="s2">
         <title>Materials and Methods</title>
         <p>During 2011 and 2012, a new field site at the Gibbs Farm (latitude: 31.4312°N and longitude: 83.5850°W) near the Coastal Plain Experiment Station, Tifton, GA was artificially inoculated with <italic>M. arenaria</italic> race 1. The susceptible peanut cultivar, ‘Georgia-10T’ (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b5">Branch and Culbreath 2011</xref>) was used to uniformly increase the peanut-specific race 1 nematode population during the growing season; whereas, hairy common vetch (<italic>Vicia villosa</italic> Roth) was used for the same purpose each winter as a susceptible cover crop.</p>
         <p>Crosses were made in the greenhouse between the RKN-susceptible parent ‘Georgia Greener’ (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b3">Branch 2007</xref>) and two advanced RKN-resistant Georgia breeding lines, GA 082524 and GA 082546 (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b6">Branch <italic>et al.</italic> 2014</xref>). These two homozygous breeding lines resulted from the same three-way cross combination ‘Georgia-02C’ (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b2">Branch 2003</xref>) × [‘Georgia-01R’ (<xref ref-type="bibr" rid="i0095-3679-43-1-49-b1">Branch 2002</xref>) × COAN].</p>
         <p>Seed of the F<sub>2</sub> and F<sub>3</sub> cross populations were space-planted 30.5-cm apart during 2013 and 2014, respectively in the newly RKN inoculated field site. Planting dates were 22 May 2013 and 23 May 2014. Recommended cultural practices with irrigation were used throughout each growing season, except that no nematicides were applied with activity against RKN. The severity of nematode galling on roots and pods was rated after digging and inverting the plants each year. All plants were dug at the same time on 21 Oct 2013 and 29 Oct 2014. The percent damage was visually estimated from 0 to 100%, with 0% representing no galls and 100% representing galling on all pods and roots. Since there can sometimes be a low level of gall formation on resistant plants, a 5% level of galling was used as a threshold in defining resistant and susceptible genotypes. Data from visual classification of segregating RKN-resistant and RKN-susceptible plants and progeny row were analyzed by a chi-square program to test goodness-of-fit of observed vs expected genetic ratios.</p>
      </sec>
      <sec id="s3">
         <title>Results and Discussion</title>
         <p>During 2013, the F<sub>2</sub> plant segregation showed an acceptable fit to both a 3:1 and 13:3, resistant to susceptible genetic ratios, respectively (<xref ref-type="table" rid="i0095-3679-43-1-49-t01">Table 1</xref>). However, the 13:3 ratio had the best fit with the lowest chi-square values and highest probability compared to the 3:1 ratio.</p>
         <table-wrap id="i0095-3679-43-1-49-t01" position="float" content-type="7col">
            <label>Table 1.</label>
            <caption>
               <p>F<sub>2</sub> plant segregation and χ<sup>2</sup> analyses for root-knot nematode (RKN) susceptible and resistant plants from two peanut cross combinations when grown in field test with high RKN populations, 2013.</p>
            </caption>
            <graphic xlink:href="i0095-3679-43-1-49-t01.png"
                     mimetype="image"
                     position="float"
                     xlink:type="simple"/>
         </table-wrap>
         <p>Because of the number of genes inherited, one of the major differences between a 3:1 vs 13:3 ratio is that the F<sub>2</sub> susceptible plants should not segregate for resistance and susceptible plants from a 3:1 ratio in the following F<sub>3</sub> generation; whereas, the F<sub>2</sub> susceptible plants would segregate from the 13:3 ratio. Both resistant and susceptible plants were found segregating within some of the F<sub>2:3</sub> susceptible progeny rows in a 1 resistant to 3 susceptible ratio.</p>
         <p>The F<sub>3</sub> progeny row segregation showed an acceptable fit to a 6 segregating to 7 non-segregating ratio for the F<sub>2:3</sub> RKN-resistant plants from the Georgia Greener x GA 082524 cross combination (<xref ref-type="table" rid="i0095-3679-43-1-49-t02">Table 2</xref>). Likewise, the F<sub>3</sub> progeny row segregation from the F<sub>2:3</sub> RKN-susceptible plants was found to have an acceptable fit to a 2 segregating to 1 non-segregating ratio as expected for a 13:3 genetic model.</p>
         <table-wrap id="i0095-3679-43-1-49-t02" position="float" content-type="6col">
            <label>Table 2.</label>
            <caption>
               <p>F<sub>3</sub> progeny row segregation and χ<sup>2</sup> analyses from F<sub>2</sub> root-knot nematode (RKN) resistant and susceptible peanut plants from the Georgia Greener × GA 082524 cross combination when grown in field test with high RKN population, 2014.</p>
            </caption>
            <graphic xlink:href="i0095-3679-43-1-49-t02.png"
                     mimetype="image"
                     position="float"
                     xlink:type="simple"/>
         </table-wrap>
         <p>The evidence from this study strongly confirms the report by <xref ref-type="bibr" rid="i0095-3679-43-1-49-b10">Church <italic>et al.</italic> (2005)</xref>. The first RKN-resistant gene should be dominant as previously reported, designated <italic>Rma<sub>1</sub>
            </italic>. However, the second RKN-resistance gene should be recessive with a proposed gene symbol, <italic>rma<sub>2</sub>
            </italic>. As expected among segregating F<sub>2</sub> plants and F<sub>2:3</sub> progeny rows in a 13:3 ratio from a two-gene model (<xref ref-type="table" rid="i0095-3679-43-1-49-t03">Table 3</xref>), either the homozygous or heterozygous dominant gene <italic>Rma<sub>1</sub> Rma<sub>1</sub>
            </italic> or <italic>Rma<sub>1</sub> rma<sub>1</sub>
            </italic>, homozygous recessive gene <italic>rma<sub>2</sub> rma<sub>2</sub>
            </italic>, or the various combinations of these two genes results in the very high level of peanut RKN resistance.</p>
         <table-wrap id="i0095-3679-43-1-49-t03" position="float" content-type="4col">
            <label>Table 3.</label>
            <caption>
               <p>An expected 13:3 ratio from a two-gene model of F<sub>2</sub> plant and F<sub>2:3</sub> progeny row segregation for peanut RKN resistance.</p>
            </caption>
            <graphic xlink:href="i0095-3679-43-1-49-t03.png"
                     mimetype="image"
                     position="float"
                     xlink:type="simple"/>
         </table-wrap>
      </sec>
   </body>
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